42-0400

Ghiroldi A, Piccoli M, Creo P, Cirillo F, Rota P, D'Imperio S, Ciconte G, Monasky MM, Micaglio E, Garatti A, Aureli M, Carsana EV, Menicanti L, Pappone C, Anastasia L
The Biochemical journal 2020 Sep 18;477(17):3401-3415

HDAC inhibitors promote pancreatic stellate cell apoptosis and relieve pancreatic fibrosis by upregulating miR-15/16 in chronic pancreatitis.

Ji T, Feng W, Zhang X, Zang K, Zhu X, Shang F
Human cell 2020 Oct;33(4):1006-1016

Effects of tacrolimus on the TGF‑β1/SMAD signaling pathway in paraquat‑exposed rat alveolar type II epithelial cells.

Ren Y, Jian X, Zhang Z, Ning Q, Kan B, Kong L
Molecular medicine reports 2020 Nov;22(5):3687-3694

Cryptotanshinone Protects Cartilage against Developing Osteoarthritis through the miR-106a-5p/GLIS3 Axis.

Ji Q, Qi D, Xu X, Xu Y, Goodman SB, Kang L, Song Q, Fan Z, Maloney WJ, Wang Y
Molecular therapy. Nucleic acids 2018 Jun 1;11:170-179

SMAD7 regulates proinflammatory and prolabor mediators in amnion and myometrium.

Lim R, Barker G, Lappas M
Biology of reproduction 2017 Aug 1;97(2):288-301

Cucurbitacin I Attenuates Cardiomyocyte Hypertrophy via Inhibition of Connective Tissue Growth Factor (CCN2) and TGF- β/Smads Signalings.

Jeong MH, Kim SJ, Kang H, Park KW, Park WJ, Yang SY, Yang DK
PloS one 2015;10(8):e0136236

Neuropilin-2 Is upregulated in lung cancer cells during TGF-β1-induced epithelial-mesenchymal transition.

Nasarre P, Gemmill RM, Potiron VA, Roche J, Lu X, Barón AE, Korch C, Garrett-Mayer E, Lagana A, Howe PH, Drabkin HA
Cancer research 2013 Dec 1;73(23):7111-21

Kruppel-like factor KLF10 targets transforming growth factor-beta1 to regulate CD4(+)CD25(-) T cells and T regulatory cells.

Cao Z, Wara AK, Icli B, Sun X, Packard RR, Esen F, Stapleton CJ, Subramaniam M, Kretschmer K, Apostolou I, von Boehmer H, Hansson GK, Spelsberg TC, Libby P, Feinberg MW
The Journal of biological chemistry 2009 Sep 11;284(37):24914-24

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of SMAD7 was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with SMAD7 Rabbit Polyclonal Antibody (Product # 42-0400) at 1 µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing Nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of SMAD7 was done on Jurkat cells treated with TNF alpha (50ng/mL, 20 minutes). Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with SMAD7 Rabbit Polyclonal Antibody (420400, red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 4 Cucurbitacin I blocks the TGF-beta/Smad signaling pathway in hypertrophic cardiomyocytes. Cell extracts (50 mug) were used for Western blot analysis of TGF-beta and Smad (Smad2, 3, and 7) protein expression levels. The expression levels of TGF-beta, Smad2, 3, and 7, and the phosphorylated forms of Smad2 and 3 (p-Smad2 and p-Smad3) were estimated by measuring band densities with NIH Image J software. GAPDH was used as a loading control. And Western blot analysis was performed in triplicate with three independent samples. Data are expressed as fold changes +- S.D. vs. control group. Significance was measured via a two-way ANOVA. * P < 0.05. Cont, control; Cu I, Cucurbitacin I.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 5 HDAC inhibition disrupts TGF-beta/Smad Signaling by miR-15 and miR-16 to protect against pancreatic fibrosis. a The protein levels and the related quantification of alpha-SMA, SMAD5 and SMAD7 in PSCs after overexpression of miR-15 or miR-16. b Immunofluorescence staining of alpha-SMA in PSCs after overexpression of miR-15 or miR-16. c The protein levels and the related quantification of alpha-SMA, SMAD5 and SMAD7 in PSCs after transfected with miR-15 inhibitor or miR-16 inhibitor with or without SAHA. All data are presented as the mean +- S.D. ( n = 3). *** P < 0.001, compared with control. Each assay was performed in triplicate