Antibody data
- Antibody Data
- Antigen structure
- References [27]
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- Validations
- Flow cytometry [1]
- Other assay [11]
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- Product number
- 56-0116-41 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD11c Monoclonal Antibody (3.9), Alexa Fluor™ 700, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The 3.9 monoclonal antibody reacts with human CD11c, the 150 kDa integrin alphaX chain. CD11c non-covalently associates with beta2 integrin to form the CD11c/CD18 heterodimer. This complex is expressed on monocytes, granulocytes, macrophages, NK, dendritic cells and subset of T and B lymphocytes. CD11c/CD18 binds to CD54, iC3b and fibrinogen and plays a role in leukocyte adhesive interactions. Applications Reported: This 3.9 antibody has been reported for use in flow cytometric analysis. Applications Tested: This 3.9 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (1 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. Alexa Fluor® 700 emits at 723 nm and can be excited with the red laser (633 nm). Most instruments will require a 685 LP mirror and 710/20 filter. Please make sure that your instrument is capable of detecting this fluorochrome. Excitation: 633-647 nm; Emission: 723 nm; Laser: Red Laser. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human
- Host
- Mouse
- Conjugate
- Near infrared dye
- Isotype
- IgG
- Antibody clone number
- 3.9
- Vial size
- 25 Tests
- Concentration
- 5 µL/Test
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references Aluminum hydroxide adjuvant diverts the uptake and trafficking of genetically detoxified pertussis toxin to lysosomes in macrophages.
Isolevuglandins disrupt PU.1-mediated C1q expression and promote autoimmunity and hypertension in systemic lupus erythematosus.
Upregulation of CD3ζ and L-selectin in antigen-specific cytotoxic T lymphocytes by eliminating myeloid-derived suppressor cells with doxorubicin to improve killing efficacy of neuroblastoma cells in vitro.
Vaccine protection against the SARS-CoV-2 Omicron variant in macaques.
Increased TNF-α Initiates Cytoplasmic Vacuolization in Whole Blood Coculture with Dengue Virus.
Low-dose Ad26.COV2.S protection against SARS-CoV-2 challenge in rhesus macaques.
IgE-Based Therapeutic Combination Enhances Antitumor Response in Preclinical Models of Pancreatic Cancer.
The long non-coding RNA Cancer Susceptibility 15 (CASC15) is induced by isocitrate dehydrogenase (IDH) mutations and maintains an immature phenotype in adult acute myeloid leukemia.
Human and Mouse Transcriptome Profiling Identifies Cross-Species Homology in Pulmonary and Lymph Node Mononuclear Phagocytes.
Unleashing Type-2 Dendritic Cells to Drive Protective Antitumor CD4(+) T Cell Immunity.
AG490 reverses phenotypic alteration of dendritic cells by bladder cancer cells.
CD226 opposes TIGIT to disrupt Tregs in melanoma.
A natural killer-dendritic cell axis defines checkpoint therapy-responsive tumor microenvironments.
Single-cell tracking of flavivirus RNA uncovers species-specific interactions with the immune system dictating disease outcome.
Resolving TYK2 locus genotype-to-phenotype differences in autoimmunity.
Effect of chronic morphine administration on circulating dendritic cells in SIV-infected rhesus macaques.
Distinct phenotype, longitudinal changes of numbers and cell-associated virus in blood dendritic cells in SIV-infected CD8-lymphocyte depleted macaques.
dNP2 is a blood-brain barrier-permeable peptide enabling ctCTLA-4 protein delivery to ameliorate experimental autoimmune encephalomyelitis.
Pharmacological targeting of miR-155 via the NEDD8-activating enzyme inhibitor MLN4924 (Pevonedistat) in FLT3-ITD acute myeloid leukemia.
Restricted dendritic cell and monocyte progenitors in human cord blood and bone marrow.
Towards programming immune tolerance through geometric manipulation of phosphatidylserine.
Breast-fed and bottle-fed infant rhesus macaques develop distinct gut microbiotas and immune systems.
TLR3-responsive, XCR1+, CD141(BDCA-3)+/CD8α+-equivalent dendritic cells uncovered in healthy and simian immunodeficiency virus-infected rhesus macaques.
Respiratory syncytial virus G protein CX3C motif impairs human airway epithelial and immune cell responses.
Pivotal role of M-DC8⁺ monocytes from viremic HIV-infected patients in TNFα overproduction in response to microbial products.
Targeting CDK1 promotes FLT3-activated acute myeloid leukemia differentiation through C/EBPα.
TNF receptor 1 genetic risk mirrors outcome of anti-TNF therapy in multiple sclerosis.
Jaldin-Fincati J, Moussaoui S, Gimenez MC, Ho CY, Lancaster CE, Botelho R, Ausar F, Brookes R, Terebiznik M
Molecular microbiology 2022 May;117(5):1173-1195
Molecular microbiology 2022 May;117(5):1173-1195
Isolevuglandins disrupt PU.1-mediated C1q expression and promote autoimmunity and hypertension in systemic lupus erythematosus.
Patrick DM, de la Visitación N, Krishnan J, Chen W, Ormseth MJ, Stein CM, Davies SS, Amarnath V, Crofford LJ, Williams JM, Zhao S, Smart CD, Dikalov S, Dikalova A, Xiao L, Van Beusecum JP, Ao M, Fogo AB, Kirabo A, Harrison DG
JCI insight 2022 Jul 8;7(13)
JCI insight 2022 Jul 8;7(13)
Upregulation of CD3ζ and L-selectin in antigen-specific cytotoxic T lymphocytes by eliminating myeloid-derived suppressor cells with doxorubicin to improve killing efficacy of neuroblastoma cells in vitro.
Xu W, Li S, Li M, Zhou H, Yang X
Journal of clinical laboratory analysis 2022 Jan;36(1):e24158
Journal of clinical laboratory analysis 2022 Jan;36(1):e24158
Vaccine protection against the SARS-CoV-2 Omicron variant in macaques.
Chandrashekar A, Yu J, McMahan K, Jacob-Dolan C, Liu J, He X, Hope D, Anioke T, Barrett J, Chung B, Hachmann NP, Lifton M, Miller J, Powers O, Sciacca M, Sellers D, Siamatu M, Surve N, VanWyk H, Wan H, Wu C, Pessaint L, Valentin D, Van Ry A, Muench J, Boursiquot M, Cook A, Velasco J, Teow E, Boon ACM, Suthar MS, Jain N, Martinot AJ, Lewis MG, Andersen H, Barouch DH
Cell 2022 Apr 28;185(9):1549-1555.e11
Cell 2022 Apr 28;185(9):1549-1555.e11
Increased TNF-α Initiates Cytoplasmic Vacuolization in Whole Blood Coculture with Dengue Virus.
Satria RD, Huang TW, Jhan MK, Shen TJ, Tseng PC, Wang YT, Yang ZY, Hsing CH, Lin CF
Journal of immunology research 2021;2021:6654617
Journal of immunology research 2021;2021:6654617
Low-dose Ad26.COV2.S protection against SARS-CoV-2 challenge in rhesus macaques.
He X, Chandrashekar A, Zahn R, Wegmann F, Yu J, Mercado NB, McMahan K, Martinot AJ, Piedra-Mora C, Beecy S, Ducat S, Chamanza R, Huber SR, van Heerden M, van der Fits L, Borducchi EN, Lifton M, Liu J, Nampanya F, Patel S, Peter L, Tostanoski LH, Pessaint L, Van Ry A, Finneyfrock B, Velasco J, Teow E, Brown R, Cook A, Andersen H, Lewis MG, Schuitemaker H, Barouch DH
Cell 2021 Jun 24;184(13):3467-3473.e11
Cell 2021 Jun 24;184(13):3467-3473.e11
IgE-Based Therapeutic Combination Enhances Antitumor Response in Preclinical Models of Pancreatic Cancer.
Markov SD, Caffrey TC, O'Connell KA, Grunkemeyer JA, Shin S, Hanson R, Patil PP, Shukla SK, Gonzalez D, Crawford AJ, Vance KE, Huang Y, Eberle KC, Radhakrishnan P, Grandgenett PM, Singh PK, Madiyalakan R, Daniels-Wells TR, Penichet ML, Nicodemus CF, Poole JA, Jaffee EM, Hollingsworth MA, Mehla K
Molecular cancer therapeutics 2021 Dec;20(12):2457-2468
Molecular cancer therapeutics 2021 Dec;20(12):2457-2468
The long non-coding RNA Cancer Susceptibility 15 (CASC15) is induced by isocitrate dehydrogenase (IDH) mutations and maintains an immature phenotype in adult acute myeloid leukemia.
Grasedieck S, Ruess C, Krowiorz K, Lux S, Pochert N, Schwarzer A, Klusmann JH, Jongen-Lavrencic M, Herold T, Bullinger L, Pollack JR, Rouhi A, Kuchenbauer F
Haematologica 2020 Sep 1;105(9):e448-453
Haematologica 2020 Sep 1;105(9):e448-453
Human and Mouse Transcriptome Profiling Identifies Cross-Species Homology in Pulmonary and Lymph Node Mononuclear Phagocytes.
Leach SM, Gibbings SL, Tewari AD, Atif SM, Vestal B, Danhorn T, Janssen WJ, Wager TD, Jakubzick CV
Cell reports 2020 Nov 3;33(5):108337
Cell reports 2020 Nov 3;33(5):108337
Unleashing Type-2 Dendritic Cells to Drive Protective Antitumor CD4(+) T Cell Immunity.
Binnewies M, Mujal AM, Pollack JL, Combes AJ, Hardison EA, Barry KC, Tsui J, Ruhland MK, Kersten K, Abushawish MA, Spasic M, Giurintano JP, Chan V, Daud AI, Ha P, Ye CJ, Roberts EW, Krummel MF
Cell 2019 Apr 18;177(3):556-571.e16
Cell 2019 Apr 18;177(3):556-571.e16
AG490 reverses phenotypic alteration of dendritic cells by bladder cancer cells.
Xiu W, Ma J, Lei T, Zhang M
Oncology letters 2018 Sep;16(3):2851-2856
Oncology letters 2018 Sep;16(3):2851-2856
CD226 opposes TIGIT to disrupt Tregs in melanoma.
Fourcade J, Sun Z, Chauvin JM, Ka M, Davar D, Pagliano O, Wang H, Saada S, Menna C, Amin R, Sander C, Kirkwood JM, Korman AJ, Zarour HM
JCI insight 2018 Jul 26;3(14)
JCI insight 2018 Jul 26;3(14)
A natural killer-dendritic cell axis defines checkpoint therapy-responsive tumor microenvironments.
Barry KC, Hsu J, Broz ML, Cueto FJ, Binnewies M, Combes AJ, Nelson AE, Loo K, Kumar R, Rosenblum MD, Alvarado MD, Wolf DM, Bogunovic D, Bhardwaj N, Daud AI, Ha PK, Ryan WR, Pollack JL, Samad B, Asthana S, Chan V, Krummel MF
Nature medicine 2018 Aug;24(8):1178-1191
Nature medicine 2018 Aug;24(8):1178-1191
Single-cell tracking of flavivirus RNA uncovers species-specific interactions with the immune system dictating disease outcome.
Douam F, Hrebikova G, Albrecht YE, Sellau J, Sharon Y, Ding Q, Ploss A
Nature communications 2017 Mar 14;8:14781
Nature communications 2017 Mar 14;8:14781
Resolving TYK2 locus genotype-to-phenotype differences in autoimmunity.
Dendrou CA, Cortes A, Shipman L, Evans HG, Attfield KE, Jostins L, Barber T, Kaur G, Kuttikkatte SB, Leach OA, Desel C, Faergeman SL, Cheeseman J, Neville MJ, Sawcer S, Compston A, Johnson AR, Everett C, Bell JI, Karpe F, Ultsch M, Eigenbrot C, McVean G, Fugger L
Science translational medicine 2016 Nov 2;8(363):363ra149
Science translational medicine 2016 Nov 2;8(363):363ra149
Effect of chronic morphine administration on circulating dendritic cells in SIV-infected rhesus macaques.
Cornwell WD, Wagner W, Lewis MG, Fan X, Rappaport J, Rogers TJ
Journal of neuroimmunology 2016 Jun 15;295-296:30-40
Journal of neuroimmunology 2016 Jun 15;295-296:30-40
Distinct phenotype, longitudinal changes of numbers and cell-associated virus in blood dendritic cells in SIV-infected CD8-lymphocyte depleted macaques.
Soulas C, Autissier PJ, Burdo TH, Piatak M Jr, Lifson JD, Williams KC
PloS one 2015;10(4):e0119764
PloS one 2015;10(4):e0119764
dNP2 is a blood-brain barrier-permeable peptide enabling ctCTLA-4 protein delivery to ameliorate experimental autoimmune encephalomyelitis.
Lim S, Kim WJ, Kim YH, Lee S, Koo JH, Lee JA, Yoon H, Kim DH, Park HJ, Kim HM, Lee HG, Yun Kim J, Lee JU, Hun Shin J, Kyun Kim L, Doh J, Kim H, Lee SK, Bothwell ALM, Suh M, Choi JM
Nature communications 2015 Sep 15;6:8244
Nature communications 2015 Sep 15;6:8244
Pharmacological targeting of miR-155 via the NEDD8-activating enzyme inhibitor MLN4924 (Pevonedistat) in FLT3-ITD acute myeloid leukemia.
Khalife J, Radomska HS, Santhanam R, Huang X, Neviani P, Saultz J, Wang H, Wu YZ, Alachkar H, Anghelina M, Dorrance A, Curfman J, Bloomfield CD, Medeiros BC, Perrotti D, Lee LJ, Lee RJ, Caligiuri MA, Pichiorri F, Croce CM, Garzon R, Guzman ML, Mendler JH, Marcucci G
Leukemia 2015 Oct;29(10):1981-92
Leukemia 2015 Oct;29(10):1981-92
Restricted dendritic cell and monocyte progenitors in human cord blood and bone marrow.
Lee J, Breton G, Oliveira TY, Zhou YJ, Aljoufi A, Puhr S, Cameron MJ, Sékaly RP, Nussenzweig MC, Liu K
The Journal of experimental medicine 2015 Mar 9;212(3):385-99
The Journal of experimental medicine 2015 Mar 9;212(3):385-99
Towards programming immune tolerance through geometric manipulation of phosphatidylserine.
Roberts RA, Eitas TK, Byrne JD, Johnson BM, Short PJ, McKinnon KP, Reisdorf S, Luft JC, DeSimone JM, Ting JP
Biomaterials 2015 Dec;72:1-10
Biomaterials 2015 Dec;72:1-10
Breast-fed and bottle-fed infant rhesus macaques develop distinct gut microbiotas and immune systems.
Ardeshir A, Narayan NR, Méndez-Lagares G, Lu D, Rauch M, Huang Y, Van Rompay KK, Lynch SV, Hartigan-O'Connor DJ
Science translational medicine 2014 Sep 3;6(252):252ra120
Science translational medicine 2014 Sep 3;6(252):252ra120
TLR3-responsive, XCR1+, CD141(BDCA-3)+/CD8α+-equivalent dendritic cells uncovered in healthy and simian immunodeficiency virus-infected rhesus macaques.
Dutertre CA, Jourdain JP, Rancez M, Amraoui S, Fossum E, Bogen B, Sanchez C, Couëdel-Courteille A, Richard Y, Dalod M, Feuillet V, Cheynier R, Hosmalin A
Journal of immunology (Baltimore, Md. : 1950) 2014 May 15;192(10):4697-708
Journal of immunology (Baltimore, Md. : 1950) 2014 May 15;192(10):4697-708
Respiratory syncytial virus G protein CX3C motif impairs human airway epithelial and immune cell responses.
Chirkova T, Boyoglu-Barnum S, Gaston KA, Malik FM, Trau SP, Oomens AG, Anderson LJ
Journal of virology 2013 Dec;87(24):13466-79
Journal of virology 2013 Dec;87(24):13466-79
Pivotal role of M-DC8⁺ monocytes from viremic HIV-infected patients in TNFα overproduction in response to microbial products.
Dutertre CA, Amraoui S, DeRosa A, Jourdain JP, Vimeux L, Goguet M, Degrelle S, Feuillet V, Liovat AS, Müller-Trutwin M, Decroix N, Deveau C, Meyer L, Goujard C, Loulergue P, Launay O, Richard Y, Hosmalin A
Blood 2012 Sep 13;120(11):2259-68
Blood 2012 Sep 13;120(11):2259-68
Targeting CDK1 promotes FLT3-activated acute myeloid leukemia differentiation through C/EBPα.
Radomska HS, Alberich-Jordà M, Will B, Gonzalez D, Delwel R, Tenen DG
The Journal of clinical investigation 2012 Aug;122(8):2955-66
The Journal of clinical investigation 2012 Aug;122(8):2955-66
TNF receptor 1 genetic risk mirrors outcome of anti-TNF therapy in multiple sclerosis.
Gregory AP, Dendrou CA, Attfield KE, Haghikia A, Xifara DK, Butter F, Poschmann G, Kaur G, Lambert L, Leach OA, Prömel S, Punwani D, Felce JH, Davis SJ, Gold R, Nielsen FC, Siegel RM, Mann M, Bell JI, McVean G, Fugger L
Nature 2012 Aug 23;488(7412):508-511
Nature 2012 Aug 23;488(7412):508-511
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Supportive validation
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- Invitrogen Antibodies (provider)
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- Staining of normal human peripheral blood cells with Mouse IgG1 K Isotype Control Alexa Fluor® 700 (Product # 56-4714-80) (blue histogram) or Anti-Human CD11c Alexa Fluor® 700 (purple histogram). Cells in the monocyte gate were used for analysis.
- Conjugate
- Near infrared dye
Supportive validation
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- Invitrogen Antibodies (provider)
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- NULL
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- Figure 4 Immune profiling in DENV-treated whole blood cells 24 h postincubation. Following DENV (MOI = 1) coculture in 100 mu l of WB ex vivo for 24 h, (a) representative flow cytometric analysis and gating of various cells obtained from five cases, performed by staining for specific cell surface markers (CD4, CD8, CD11c, CD14, CD16, CD19, CD25, CD56, CD62L, and HLA-DR), in the DENV-infected and mock groups showed (b) the changes in the expression of specific immune cell populations as noted. (c) The results are shown as a percentage of the mean +- SD obtained from five cases. * p < 0.05, ** p < 0.01, and *** p < 0.001, compared to the mock group. R: region; WBC: white blood cell; bri: bright.
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- Figure 1. Gating Strategy to Sort Human and Mouse Mononuclear Phagocytes for RNA Isolation (A and B) Fluorescence-activated cell sorting (FACS) gating strategy used to isolate human (A) or mouse (B) mononuclear phagocytes (MPs, labeled in red) from the indicated tissues. Before sorting, cell suspensions were magnetically enriched as indicated and gated to select Live; Single; CD45 + ; Lineage - cells as shown in Figures S1 and S2 . Arrows indicate where further analysis is performed on the specified subpopulation. The data presented are representative of 3-6 replicates per sort. (C) Table illustrating cell surface marker expression by MP subsets in human (top) and mouse (bottom). - or lo signifies no or little expression, + or ++ indicates expression or strong expression, -/+ indicates heterogeneous expression of marker proteins.
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- FIGURE 1 Extraction, identification, and purification of myeloid-derived suppressor cell and cultivation of dendritic cell. (A) Cells were extracted from the bone marrow of BALB/c mice and stained by monoclonal antibodies. Under flow cytometry, the expressive rate of Gr-1 + MDSC, CD11b + MDSC, CD11c + MDSC, CD80 + MDSC, F4/80 + MDSC, and MHC-II + MDSC were 70.4%, 3.5%, 4.8%, 1.2%, 0.3%, 2.1% respectively. (B) The expressive rate of Gr-1 + CD11b + MDSC was 22.6%. (C) After MACS by CD11b magnetic bead, purification of Gr-1 + CD11b + MDSC reached 84.6%. (D) Most non-antigen-loaded dendritic cells grew adherently, with different sizes, star or spindle shape, and stretching tubers, but some of the cells seemed to have adopted a half-adherent state with rough surface. The expressive rates of CD11c, CD86, and MHC-II on DCs were 10.9%, 3.8%, and 27.9%, respectively, by flow cytometry. (E) On the 7th day, DCs were stimulated and activated by tumor antigens. DCs in the half-adherent state increased obviously with radial spikes and bigger shape. The expressive rates of CD11c, CD86, and MHC-II were 74.8%, 50.3%, and 49.8%, respectively, by flow cytometry. IgG FITC, a homotypic control antibody, was used to set the gate strategy. Scale bar = 100 mumol/liter. MDSC, myeloid-derived suppressor cell; MACS, magnetic-activated cell sorting; DC, dendritic cell; CD, cluster of differentiation
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- 5 FIGURE Impact of particle size on aluminum adjuvant internalization. (a) U-937 cells were incubated with AlOOH-lumo (control) or AlOOH-lumo previously subjected to freeze/thaw cycles (freeze/thaw) to increase the size of the particles, as described in experimental procedures. The cells were incubated at 37degC for 4 h and subsequently processed for flow cytometry analysis. Differentiated macrophages were classified using anti-CD11c antibodies. Cell nuclei were labeled with DAPI. Cell populations encircled by a dotted line have not internalized AlOOH-Lumo and cells that have internalized AlOOH-Lumo are encircled by a solid line. (b) U-937 cells were incubated with control or freeze/thaw AlOOH-lumo preparations as described above and subsequently fixed and F-Actin stained with blue phalloidin (pseudocolored gray). Red arrowheads in the right panel point to small, internalized adjuvant particles. Spinning disk confocal images represent a merge of z-stacks. Images are representative of three independent trials. Fifty cells per trial per condition were analyzed. Scale bars, 5 mum. (c) U-937 cells were incubated with control or freeze/thaw AlOOH-lumo preparations and processed as described in (b). The percentage of cells with intracellular AlOOH particles were calculated for three independent trials and the mean represented in the bar graph. * p
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- IsoLG adducts are enriched in monocytes of patients with SLE. ( A ) Representative FACS plots displaying isoLG adduct containing CD11c + PBMCs from a representative control and patient with SLE. Representative histograms displaying the distribution of isoLG adducts in ( B ) CD11c + and ( C ) CD11c + CD86 + cells. Quantitation of IsoLG adduct-containing cells as a percentage of ( D ) CD11c + , ( E ) CD11c + CD86 + , and ( F ) CD14 + cells. For B - F data were analyzed using 1-tailed Student's t test or Mann-Whitney U test ( n = 10-11, *P < 0.05). ( G ) Stable isotope dilution multiple reaction monitoring for mass spectrometry analysis of isoLG-lysine-lactam adduct in DCs. Representative liquid chromatography/mass spectrometry chromatographs from a representative patient. The top row shows multiple reaction monitoring chromatographs for isoLG lysine lactam in samples, while the bottom row shows multiple reaction monitoring chromatograph for [13C615N2] internal standard for the same samples. cps, counts per second; Rt, retention time. ( H ) Quantitation of isoLG-lysine in monocytes from a subset of SLE patients and controls. ( I ) Monocytes from SLE patients and controls were sorted. Superoxide was detected using HPLC to monitor conversion of dihydroethidium to the superoxide oxidation adduct 2-hydroxyethidium (2-HO-ET) and ethidium. ( J ) Quantitation of 2-HO-ET from SLE patients and controls. For H and J , comparisons were made with a 1-tailed Student's t test ( n = 4-6, *P
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- Figure 2 Protein delivery efficiency of dNP2 in primary mouse and human immune cells. ( a , b ) Mouse primary splenocytes were isolated from 6-week-old female C57BL/6 mice and the cells were incubated with 5 muM EGFP, TAT- and dNP2-EGFP for 2 h. Intracellular fluorescence was analysed by flow cytometry and the data are represented as dot plots or mean fluorescence intensity (MFI) of the cells. ( c , d ) Human PBMCs were isolated from healthy donor blood and the cells were incubated with 5 muM EGFP, TAT-, dNP2-EGFP for 2 h. The data were analysed as described above. ( e ) Total splenocytes were incubated with 1 muM EGFP, TAT-, and dNP2-EGFP for 2 h. Cells were gated using markers specific for CD4 T cells (CD4 + ), B cells (CD19 + ), macrophages (CD11c lo CD11b hi F480 + ) and DCs (CD11c hi MHCII hi ). The EGFP signal in each cell population was then analysed by flow cytometric analysis. The relative MFI value was normalization to PBS treated cells. The red line indicates relative MFI of PBS-treated cells. ( f ) Total PBMCs were incubated with 1 muM EGFP, TAT-, and dNP2-EGFP for 2 h. Cells were gated with markers specific for CD4 T cells (CD4 + ), B cells (CD19 + ), macrophages (CD11b + ) and DCs (CD11c + ) and the data were then analysed as described above. ( g ) Time-lapse images of mouse CD4 T cells incubated with 1 muM EGFP, TAT- and dNP2-EGFP were acquired for 2 h (Scale bar, 15 mum) and ( h ) the average fluorescence intensities of 10 cells from each sample were calculate
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- Figure 2. Percentage of purified DC samples in all experiments. DCs were labeled with the Non-DC Depletion Cocktail for negative selection. Next, DCs were labeled with DC Enrichment Cocktail for positive selection. The expression of CD11c and CD86 was tested by flow cytometery. The percentages of purified DC are presented in the top of each panel. DCs, dendritic cells; CD, cluster of differentiation.
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- Figure 1. CASC15 -KO promotes the differentiation of acute myeloid leukemia cells. (A) Apoptosis in CASC15 -KO and empty vector-transduced (control) OCI-AML5 cell lines after 24 h of depletion of granulocyte-macrophage colony-stimulating factor (annexin-FITC/Sytox blue flow cytometry). (B) Expression of SOX4 during in vitro differentiation of CASC15 -KO and control OCI-AML5 cell lines. All cells were treated with 0.1 mM all- trans retinoic acid (ATRA) and 1 nM 12-O-tetradecanoylphorbol-13-acetate (TPA) over 72 h in three independent experiments. Total RNA was extracted before, after 24 h and after 72 h of treatment, DNase-digested and transcribed to cDNA. A quantitative real-time polymerase chain reaction (qRT-PCR) was performed using SYBR green chemistry with subsequent melting curve analysis in technical triplicates. The 2-ddCt was calculated relative to the pre-determined housekeeping gene encoding succinate dehydrogenase complex subunit C ( SDHC ). (C) Baseline expression of the monocyte/macrophage markers CD11b (integrin subunit alpha M, ITGAM), CD11c (integrin subunit alpha X, ITGAX), and CD14, the granulocyte marker CD15 (fucosyltransferase 4, FUT4), and the general myeloid marker CD13 (aminopeptidase N, APN) in CASC15 -KO and control cells. The percentages of positive cells, quantified by flow cytometry after 72 h, are shown. (D-F) Growth rate and CD11c myeloid cell surface marker expression of CASC15 and control cell lines during drug-induced in vitro differentiation
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