12-4877-41

antibody from Invitrogen Antibodies

Targeting: EOMES TBR2

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

12-4877-41

Provider

Invitrogen Antibodies

Product name

EOMES Monoclonal Antibody (WD1928), PE, eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: This WD1928 antibody recognizes Eomesodermin (Eomes), also known as T-box brain 2 (TBR2). Eomes is a T-box transcription factor that is highly homologous to T-bet, which is essential during trophoblast development and gastrulation in most vertebrates. In the immune system, Eomes controls the differentiation of effector and memory CD8+ T cells, as well as natural killer (NK) cells. Expression of Eomes in these cells correlates with high expression of CD122, the common beta-chain of the IL-2R and IL-15R. Applications Reported: This WD1928 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This WD1928 antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of normal human peripheral blood cells using the Foxp3/Transcription Factor Buffer Set (Product # 00-5523-00) and protocol. Please refer to Best Protocols: Protocol B: One step protocol for (nuclear) intracellular proteins. This can be used at 5 µL (0.25 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. Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser. Filtration: 0.2 µm post-manufacturing filtered.

Reactivity

Human, Porcine

Host

Mouse

Conjugate

Yellow dye

Isotype

IgG

Antibody clone number

WD1928

Vial size

25 Tests

Concentration

5 µL/Test

Storage

4° C, store in dark, DO NOT FREEZE!

References

Submitted references

Transcriptional profiling of human Vδ1 T cells reveals a pathogen-driven adaptive differentiation program.

McMurray JL, von Borstel A, Taher TE, Syrimi E, Taylor GS, Sharif M, Rossjohn J, Remmerswaal EBM, Bemelman FJ, Vieira Braga FA, Chen X, Teichmann SA, Mohammed F, Berry AA, Lyke KE, Williamson KC, Stubbington MJT, Davey MS, Willcox CR, Willcox BE
Cell reports 2022 May 24;39(8):110858

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A single-cell map of dynamic chromatin landscapes of immune cells in renal cell carcinoma.

Kourtis N, Wang Q, Wang B, Oswald E, Adler C, Cherravuru S, Malahias E, Zhang L, Golubov J, Wei Q, Lemus S, Ni M, Ding Y, Wei Y, Atwal GS, Thurston G, Macdonald LE, Murphy AJ, Dhanik A, Sleeman MA, Tykodi SS, Skokos D
Nature cancer 2022 Jul;3(7):885-898

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Homeostatic Function and Inflammatory Activation of Ileal CD8(+) Tissue-Resident T Cells Is Dependent on Mucosal Location.

Lutter L, Roosenboom B, Brand EC, Ter Linde JJ, Oldenburg B, van Lochem EG, Horjus Talabur Horje CS, van Wijk F
Cellular and molecular gastroenterology and hepatology 2021;12(5):1567-1581

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Norovirus-Specific CD8(+) T Cell Responses in Human Blood and Tissues.

Pattekar A, Mayer LS, Lau CW, Liu C, Palko O, Bewtra M, Consortium H, Lindesmith LC, Brewer-Jensen PD, Baric RS, Betts MR, Naji A, Wherry EJ, Tomov VT
Cellular and molecular gastroenterology and hepatology 2021;11(5):1267-1289

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Culture, expansion, and flow-cytometry-based functional analysis of pteropid bat MR1-restricted unconventional T cells.

Sia WR, Hey YY, Foo R, Wang LF, Leeansyah E
STAR protocols 2021 Jun 18;2(2):100487

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Complement opsonization of HIV affects primary infection of human colorectal mucosa and subsequent activation of T cells.

Bhattacharya P, Ellegård R, Khalid M, Svanberg C, Govender M, Keita ÅV, Söderholm JD, Myrelid P, Shankar EM, Nyström S, Larsson M
eLife 2020 Sep 2;9

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CD19-CAR-T Cells Bearing a KIR/PD-1-Based Inhibitory CAR Eradicate CD19(+)HLA-C1(-) Malignant B Cells While Sparing CD19(+)HLA-C1(+) Healthy B Cells.

Tao L, Farooq MA, Gao Y, Zhang L, Niu C, Ajmal I, Zhou Y, He C, Zhao G, Yao J, Liu M, Jiang W
Cancers 2020 Sep 13;12(9)

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Cell-type-specific 3D epigenomes in the developing human cortex.

Song M, Pebworth MP, Yang X, Abnousi A, Fan C, Wen J, Rosen JD, Choudhary MNK, Cui X, Jones IR, Bergenholtz S, Eze UC, Juric I, Li B, Maliskova L, Lee J, Liu W, Pollen AA, Li Y, Wang T, Hu M, Kriegstein AR, Shen Y
Nature 2020 Nov;587(7835):644-649

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Single-cell transcriptomics reveals regulators underlying immune cell diversity and immune subtypes associated with prognosis in nasopharyngeal carcinoma.

Chen YP, Yin JH, Li WF, Li HJ, Chen DP, Zhang CJ, Lv JW, Wang YQ, Li XM, Li JY, Zhang PP, Li YQ, He QM, Yang XJ, Lei Y, Tang LL, Zhou GQ, Mao YP, Wei C, Xiong KX, Zhang HB, Zhu SD, Hou Y, Sun Y, Dean M, Amit I, Wu K, Kuang DM, Li GB, Liu N, Ma J
Cell research 2020 Nov;30(11):1024-1042

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The TLR9 ligand CpG ODN 2006 is a poor adjuvant for the induction of de novo CD8(+) T-cell responses in vitro.

Papagno L, Kuse N, Lissina A, Gostick E, Price DA, Appay V, Nicoli F
Scientific reports 2020 Jul 15;10(1):11620

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The CD38/NAD/SIRTUIN1/EZH2 Axis Mitigates Cytotoxic CD8 T Cell Function and Identifies Patients with SLE Prone to Infections.

Katsuyama E, Suarez-Fueyo A, Bradley SJ, Mizui M, Marin AV, Mulki L, Krishfield S, Malavasi F, Yoon J, Sui SJH, Kyttaris VC, Tsokos GC
Cell reports 2020 Jan 7;30(1):112-123.e4

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C reactive protein impairs adaptive immunity in immune cells of patients with melanoma.

Yoshida T, Ichikawa J, Giuroiu I, Laino AS, Hao Y, Krogsgaard M, Vassallo M, Woods DM, Stephen Hodi F, Weber J
Journal for immunotherapy of cancer 2020 Apr;8(1)

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Tissue-resident Eomes(+) NK cells are the major innate lymphoid cell population in human infant intestine.

Sagebiel AF, Steinert F, Lunemann S, Körner C, Schreurs RRCE, Altfeld M, Perez D, Reinshagen K, Bunders MJ
Nature communications 2019 Feb 28;10(1):975

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Unique transcriptional and protein-expression signature in human lung tissue-resident NK cells.

Marquardt N, Kekäläinen E, Chen P, Lourda M, Wilson JN, Scharenberg M, Bergman P, Al-Ameri M, Hård J, Mold JE, Ljunggren HG, Michaëlsson J
Nature communications 2019 Aug 26;10(1):3841

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Mass cytometry reveals innate lymphoid cell differentiation pathways in the human fetal intestine.

Li N, van Unen V, Höllt T, Thompson A, van Bergen J, Pezzotti N, Eisemann E, Vilanova A, Chuva de Sousa Lopes SM, Lelieveldt BPF, Koning F
The Journal of experimental medicine 2018 May 7;215(5):1383-1396

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Systemic Human ILC Precursors Provide a Substrate for Tissue ILC Differentiation.

Lim AI, Li Y, Lopez-Lastra S, Stadhouders R, Paul F, Casrouge A, Serafini N, Puel A, Bustamante J, Surace L, Masse-Ranson G, David E, Strick-Marchand H, Le Bourhis L, Cocchi R, Topazio D, Graziano P, Muscarella LA, Rogge L, Norel X, Sallenave JM, Allez M, Graf T, Hendriks RW, Casanova JL, Amit I, Yssel H, Di Santo JP
Cell 2017 Mar 9;168(6):1086-1100.e10

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Human Th1 cells that express CD300a are polyfunctional and after stimulation up-regulate the T-box transcription factor eomesodermin.

Narayanan S, Silva R, Peruzzi G, Alvarez Y, Simhadri VR, Debell K, Coligan JE, Borrego F
PloS one 2010 May 13;5(5):e10636

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Intracellular staining of normal human peripheral blood cells with Anti-Human CD8 APC (Product # 17-0087-42) Mouse IgG1 K Isotype Control PE (left) or Anti-Human Eomes PE (right) using the Foxp3/Transcription Factor Buffer Set (Product # 00-5523-00) and protocol. Cells in the lymphocyte gate were used for analysis.

Conjugate

Yellow dye

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 1. Experimental design and features of 3D epigenomes during human corticogenesis. ( a ) Schematic of the sorting strategy. Microdissected GZ and CP samples were dissociated into single cells prior to being fixed, stained with antibodies for PAX6, SOX2, EOMES, and SATB2, and sorted using FACS. ( b ) Heatmap displaying the expression of key marker genes for each cell type. ( c ) WashU Epigenome Browser snapshot displaying a region (chr17: 72,970,000-73,330,000) with interactions linked to SSTR2 expression in IPCs. ( d ) Bar graph of interaction counts for each cell type, with the proportions of anchor to anchor (red) and anchor to non-anchor (blue) interactions highlighted. ( e ) Cumulative distribution function (CDF) plots of interaction distances for each cell type. ( f ) Histogram displaying the numbers of interactions for interacting promoters across all cell types.

Conjugate

Yellow dye

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Extended Data Figure 1. Representative contour plots depicting FACS gating strategy. ( a ) Cells were separated from debris of various sizes based on the forward scatter area (FSC-A) and side scatter area (SSC-A). Specifically, they were passed through two singlet gates using the width and height metrics of the ( b ) side scatter (SSC-H versus SSC-W) and ( c ) forward scatter (FSC-H versus FSC-W). ( d ) SOX2+, and SOX2-, and intermediate progenitor (IPC) populations were isolated by gating on EOMES-PE-Cy7 and SOX2-PerCP-Cy5.5 staining. ( e ) Radial glia (RG) and interneurons (iNs) were isolated based on high PAX6/high SOX2 and medium SOX2/low PAX6 staining, respectively. ( f ) Excitatory neurons (eNs) were isolated from the SOX2- population by gating on SATB2-Alexa Fluor 647 staining.

Conjugate

Yellow dye

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 13 Pa MR1T cell antigen-specific proliferation assay (A) Representative FACS plots showing frequencies of Pa MR1T cells (defined as CD3 + hMR1-5-OP-RU + ) and CTV dilution on day 0 and day 5 of culture with 5-OP-RU from a single Pa donor. (B) Representative FACS plots showing the expression levels of different transcription factors (PLZF, T-bet, Eomes, and RORgammat) and Perforin against CTV at the two different timepoints. Figure adapted and reprinted with permission from ().

Conjugate

Yellow dye

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 6. HIV exposure modulated transcriptional regulation of mucosal T cells. Mucosal immune cells isolated from the colorectal tissue biopsies by enzymatic digestion were exposed to HIV-1 BaL (250 ng/mL), either free (F-HIV), complement-opsonized (C-HIV), or virions opsonized by a cocktail of complement and antibodies (CI-HIV) or mock-treated, by spinning the cultures. The effect HIV exposure had on mucosal T cells phenotype was assessed after 4 days. ( A-D ) The immune cells were stained with CD3, CD4, CD8, Tbet, EOMES, PD-1, LAG3, TIM3, and CD160 mAbs. ( A ) Percentage of unexposed, F-HIV, C-HIV, or CI-HIV exposed Tbet+CD4+ T cells expressing PD-1, TIM3, LAG3 or CD160. viSNE plot of PD-1+Tbet+CD4+ was constructed on CD4 T cells, data presented as dot plots with colored channels. Data from a donor shows CD4 T cells, subjected to tSNE algorithm, which provides cells with a unique coordinate according to its expression of PD-1+T-bet+ parameters, displayed on a two-dimensional plot (tSNE1 versus tSNE2). The heat gradient (blue to red) indicates expression level of Tbet+PD-1+ on CD4 following exposure to different conditions of viruses. ( B ) Percentage of unexposed, F-HIV, C-HIV, or CI-HIV exposed Tbet+EOMES+ CD4+ T cells expressing PD-1, TIM3, LAG3 or CD160 were assessed by flow cytometry. Flow cytometry Zebra plots for Tbet+EOMES+ CD4+ T cells expressing PD-1. ( C ) Percentage of Tbet+CD8+ T cells expressing PD-1, TIM3, LAG3 or CD160. viSNE plot of PD-1+Tbet+CD8+ was const

Conjugate

Yellow dye

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 7 Phenotypic characterization of NoV-specific CD8 + T cells. ( A ) PBMCs or ( B ) LPMCs were stained with tetramers and a panel of antibodies against memory, homing, exhaustion, proliferation, and cytotoxicity markers. Total live CD8 + T cells are shown in grey with Tet + cells overlayed in red . Representative of 3 independent experiments.

Conjugate

Yellow dye

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 3 Subset defining genes of lamina propria CD8 + CD69 + CD103 - and CD8 + CD69 + CD103 + T cells. ( A ) Volcano plot of the expressed genes, with a nominal P value 0.25 and -log10 P > 10 x 10 -2.5 , green indicates genes with a log2FC >0.25 and -log10 P < 10 x 10 -2.5 . ( B ) Representative flowcytometric dotplots, including Fluorescence Minus One (FMO) control, of Itgb2, GzmK, KLRG1, and EOMES (upper row) and quantification of the respective marker (lower row) comparing lamina propria CD8 + CD69 + CD103 + and CD103 - T cells in healthy control subjects (n = 6-7; circles), CD patients from inflamed (diamonds) and noninflamed (triangles) ileum (paired, n = 4-6). Bars represent median and interquartile range. Comparison was performed with a paired 1-tailed t test. NS, not significant.

Conjugate

Yellow dye

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

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Experimental details

Fig. 4 NF-kappaB drives a pro-apoptotic program in late dysfunctional CD8 + T cells infiltrating ccRCC. a , UMAP projection of scATAC-seq profiles colored by chromVAR TF motif bias-corrected deviations of the indicated TFs (NFKB1/2 and RELA/B were significantly enriched in C11_CD8 compared with the other CD8 T cell clusters; two-sided Wilcoxon rank-sum test P < 2.2 x 10 -16 , n = 18,736 cells). b , Violin plots of gene activity scores of the indicated NF-kappaB transcriptional targets for effector/nondysfunctional (C4,5,7_CD8) and dysfunctional (C8,9,10,11_CD8) T cell clusters. Pairwise comparisons of gene activity scores for the indicated gene between specified T cell clusters were determined using a two-sided Wilcoxon rank-sum test. The resulting P values underwent multi-test correction with the FDR method (**adjusted P < 0.01, ****adjusted- P < 0.0001; raw adjusted P values are listed in the source data; for clusters 4, 5, 7, 8, 9, 10, 11, n = 1,810, 1,821, 873, 924, 1,775, 2,440 and 1,613 cells, respectively). c , Violin plots showing gene expression levels of the indicated NF-kappaB transcriptional targets in CD4 + and CD8 + clusters, clustered as indicated in Fig. 3a (for clusters 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 and 14, n = 8,677, 7,423, 5,390, 5,272, 4,469, 4,160, 3,974, 3,689, 1,080, 890, 751, 669, 618 and 328 cells, respectively). d , Flow cytometry analysis of tumor-infiltrating CD8 + T cells. UMAP projection of expression of selected T cell markers in CD8

Conjugate

Yellow dye

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 2 Human CD69 + CD16 - lung NK cell subsets have unique characteristics. a Representative overlays and b summary of data of expression of NKG2A and CD57 on CD16 - human lung CD69 - CD49a - CD103 - ( n = 20 for both), CD69sp ( n = 20 for both), CD69 + CD49a + CD103 - ( n = 14 and n = 13, respectively), and CD69 + CD49a + CD103 + ( n = 17 and n = 15, respectively) NK cells. CD56 dim CD16 + NK cells are shown for comparison ( n = 20 and n = 19, respectively). Numbers in a indicate %NKG2A + and %CD57 + NK cells, respectively. c Representative overlays and d summary of mean fluorescence intensity (MFI) of CD56 ( n = 18 for CD16 - CD69 - CD49a - CD103 - NKG2A + CD57 - , n = 21 for CD69 + CD49a - CD103 - CD16 - , n = 16 for CD69 + CD49a + CD103 - CD16 - , n = 19 for CD69 + CD49a + CD103 + CD16 - , and n = 20 for CD56 dim CD16 + ), CXCR3 ( n = 3) and CXCR6 ( n = 7) on human lung CD69 - CD49a - CD103 - NKG2A + CD57 - , CD69sp, CD69 + CD49a + CD103 - , and CD69 + CD49a + CD103 + NK cells within the CD16 - subset. e Representative overlay and f summary of MFI of perforin expression in CD16 - CD69 - CD49a - CD103 - NKG2A + CD57 - , CD69spCD16 - , CD69 + CD49a + CD103 - CD16 - , CD69 + CD49a + CD103 + CD16 - , and CD69 - CD49a - CD103 - CD16 + NK cells in human lung ( n = 5). CD127 + CD161 + cells from lung tissue are shown as a comparison ( n = 5). g Representative overlays and h summary of MFI of Eomes (left panel) (CD16 - CD69

Conjugate

Yellow dye

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

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Experimental details

Fig. 5 High expression of Eomes in infant intestinal NK cells. a viSNE plots of combined flow cytometric data visualizing Eomes expression of infant and adult NK cells from epithelium (EP) and lamina propria (LP) intestinal tissues. Cell density of clusters is shown in first row. Expression of Eomes is shown by color coding in relative intensity in second row. viSNE plots have been calculated from concatenated FCS files gated on NK cells (infant samples N = 7, adult samples N = 5, iterations = 7500 perplexity = 100, KL divergence = 2.62). b Frequencies of EP and LP-derived infant (white circles) and adult (dark circles) Eomes + NK cells. c Representative flow cytometric plots showing co-expression of Eomes and tissue-residency markers (CD103, CD49a, CD69) in NK cells from EP and LP of infant and adult intestines. d Frequencies of Eomes + NK cells within CD103 + , CD49a + , or CD69 + NK cell populations in EP and LP of infants (white circles) and adults (dark circles) (infant samples N = 8 (EP and LP), adult samples N = 8 (EP), and N = 10 (LP)). Median frequencies indicated by red lines. Error bars define interquartile ranges between 75th and 25th percentiles. Statistical comparisons are Mann-Whitney U comparisons ( b ) and Wilcoxon matched-pairs signed rank tests ( d ). Asterisks represent the following p -values: * p < 0.05; ** p < 0.01; and *** p < 0.001

Conjugate

Yellow dye

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 6 Infant intestinal NK cells contain high levels of cytotoxic granules. a viSNE plots of combined flow cytometric data visualizing Eomes, perforin, granzyme B, and KIR expression by epithelial (EP) and lamina propria-derived (LP) infant and adult NK cells. Expression of Eomes, KIR, perforin, and granzyme B (GrzB) is shown by color coding in relative intensity. viSNE plots have been calculated from concatenated FCS files gated on NK cells (infant samples N = 7, adult samples N = 5, iterations = 7500 perplexity = 100, KL divergence = 2.62). b Frequencies of perforin + and GrzB + NK cells in infants (white circles) and adults (dark circles) (EP infant samples ( N = 8), LP infant samples ( N = 7), EP adult samples perforin expression ( N = 9), GrzB expression ( N = 8), LP adult samples perforin expression ( N = 9), and GrzB expression ( N = 8)). c Frequencies of perforin + and granzyme B + cells within CD103 + , CD49a + , or CD69 + NK cell populations in infant (white circles) and adult intestines (dark circles) (EP infant samples ( N = 8), LP infant samples ( N = 7), EP adult samples perforin expression ( N = 9), and GrzB expression ( N = 8), LP adult samples perforin expression ( N = 9), and GrzB expression ( N = 8)). d Frequencies of LP-derived CD107a + , IFN-g + , and TNF-alpha + NK cells in infant (white circles) and adult intestines (dark circles). Cells were stimulated for 6 h with phorbol 12-myristate 13-acetate (PMA) and ionomycin (infant samples N = 5, adult sample

Conjugate

Yellow dye

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 3 Characteristics of iKP-19-CAR-T cells and CD19-CAR-T cells. iKP-19-CAR-T cells or CD19-CAR-T cells were cultured for 10 days in X-VIVO media supplemented with 100 U/mL IL-2. ( A ) The expression of T cell differentiation markers in CAR-T cells was analyzed by flow cytometry using PE/Cy7-anti-human CCR7 antibody, PE-anti-human-CD45RO antibody and PE-anti-human GzmB antibody ( n = 4 different donors). ( B ) The frequency of naive (T N ; CCR7 + CD45RA + ), T CM (CCR7 + CD45RA - ), effector memory (T EM ; CCR7 - CD45RA - ) or effector (T E ; CCR7 - CD45RA + ) T cells were analyzed by flow cytometry using PE/Cy7-anti-human CCR7 antibody and FITC-anti-human CD45RA antibody ( n = 4 different donors). ( C ) The expression of transcription factor Eomes in CAR-T cells was analyzed by flow cytometry using FITC-anti-human Eomes antibody ( n = 4 different donors). ( D ) The expression of transcription factor T-bet in CAR-T cells was analyzed by flow cytometry using PE-anti-human T-bet antibody ( n = 4 different donors). ( E ) The expression of T cell exhaustion marker PD-1 in CAR-T cells was analyzed by flow cytometry using FITC-anti-human PD-1 antibody ( n = 4 different donors). All experiments were performed in triplicate manner using PBMCs from each donor and MFI or percentage was statistically analyzed. * p < 0.05, ** p < 0.01, *** p < 0.001. Error bars represent +- SD.

Conjugate

Yellow dye

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 4. CD8CD38 high T Cells Express Decreased Levels of EOMES, RUNX3, and T-bet (A) Scheme showing the transcription factors regulating various cytotoxic molecules. (B) Representative histograms showing EOMES, RUNX3, and T-bet in CD8CD38 low and CD38 high T cells from a healthy donor by flow cytometry. (C) MFI from EOMES, RUNX3, and T-bet in CD8CD38 low and CD8CD38 high T cells from by flow cytometry (healthy subjects >=3; paired t test). (D) Percentage of EOMES, RUNX3, and T-bet in CD8CD38 low and CD8CD38 high T cells by flow cytometry (SLE = 6; paired t test). (E) mRNA levels of EOMES, RUNX3, and T-bet (TBX21) in CD8CD38 low and CD8CD38 high T cells by flow cytometry (normal T cells = 10; Kolmogorov-Smirnov test). Cells were sorted by FACS aria. In all figures, average data are represented as mean +- SD.

Conjugate

Yellow dye

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 6. Inhibition of Sirtuin1/EZH2 Axis Restores CD8 Cytotoxicity (A) MFI from EOMES, RUNX3, and T-bet in CD8 T cells pretreated with 50 muM of EX527 (SIRT1 inhibitor) overnight by flow cytometry (healthy subjects = 7; paired t test). (B) MFI from EOMES and RUNX3 in TALL104 CD8 T cells pretreated with 1 muM of GSK126 (EZH2 inhibitor) overnight by flow cytometry (n = 4, 3 independent experiments; paired t test). (C) MFI from RUNX3 in CD8 T cells in patients with SLE pretreated with 1 muM of GSK126 overnight by flow cytometry (SLE = 4; paired t test). (D and E) Percentage of IFN-gamma and GZMB in CD8CD38 low or CD8CD38 high T cells from healthy controls (D) and from patients with SLE (E) treated with 1 muM of GSK126 overnight by flow cytometry (healthy subjects = 5, shown in closed circle, SLE = 6, shown in opened circle; paired t test). (F and G) Degranulation (%CD107a) of CD8CD38 low or CD8CD38 high T cells from healthy subjects (F) and from patients with SLE (G) treated with GSK126 overnight and stimulated with plate-coated CD3/CD28 antibodies for 5 h with GSK126 at the indicated concentration (n = 4, one-way ANOVA with multiple comparisons).

Conjugate

Yellow dye