Sb431542 s4317

Manufactured by Merck Group

Sourced in Macao, United States

SB431542 (S4317) is a transforming growth factor-beta (TGF-β) receptor inhibitor used in cell culture and research applications. It functions by blocking the activation of the TGF-β signaling pathway. This product is intended for research use only and its detailed applications should be evaluated by the user.

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4 protocols using sb431542 s4317

Antibodies and Reagents for Cell Culture

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The primary antibodies used in this study are listed in the supplementary material. The secondary antibodies were purchased from Zhongshan Golden Bridge Biotechnology Co., Ltd. (Beijing, China). SB431542 (S4317) was purchased from Sigma-Aldrich (St. Louis, MO), and SB431542 (S1067) was purchased from Selleckchem (Houston, TX). Matrigel was purchased from BD Biosciences (NY, USA).

Gong W., Sun B., Zhao X., Zhang D., Sun J., Liu T., Gu Q., Dong X., Liu F., Wang Y., Lin X, & Li Y. (2016). Nodal signaling promotes vasculogenic mimicry formation in breast cancer via the Smad2/3 pathway. Oncotarget, 7(43), 70152-70167.

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Epithelial-Mesenchymal Transition Antibody Assay

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Antibodies against Slug (SC‐10436), Twist (SC‐15393) and β‐actin (sc‐47778) were purchased from Santa Cruz (CA, USA); those against cleaved caspase‐3 (9661), cleaved PARP (9541), TGF‐β (3711), Smad2 (5339), p‐Smad2 (3108), Smad3 (9523) and p‐Smad3 (9520) were from Cell Signaling Technology (MA, USA); and those against N‐cadherin (610921) and E‐cadherin (610182) were from BD, NJ, USA. Anti–HAS2 (ab140671), anti–Vimentin (3634‐100) and anti–zeb1 (HPA027524) were from Abcam (MA, USA), Biovision (CA, USA) and Sigma (MO, USA), respectively. Hyaluronan (GLR001, low molecular weight; GLR004, middle molecular weight; GLR002, high molecular weight) was purchased from R&D Systems (MN, USA). 4‐Methylumbelliferone (M1381) and SB431542 (S4317) was purchased from Sigma (MO, USA).

Kim Y., Lee S.B., Shim S., Kim A., Park J., Jang W., Lee S., Myung J.K., Park S., Lee S, & Kim M. (2019). Hyaluronic acid synthase 2 promotes malignant phenotypes of colorectal cancer cells through transforming growth factor beta signaling. Cancer Science, 110(7), 2226-2236.

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Signaling Pathway Antibody Validation

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Antibodies used in this study were commercially obtained as the following: anti‐PAI-1 (C-9) antibody from Santa Cruz Biotech; anti‐p21 Waf1/Cip1 (2947 S), anti-p15 INK4B (4822 S), anti-p57/Kip2 (2557 S), anti-p-Smad2 (Ser465/467) /Smad3 (Ser423/425) (8828 S), anti-Smad2/3 (8685 S), anti-Smad4 (38454 S), anti‐HA (3724), anti-Myc (2276 S), anti-Caspase-3 (2662 S), anti-PARP (9532 S), anti-p-Tyrosine/P-Tyr-100 (9411 S), anti-ABL1 (2862 S), anti-Ac-Histone H3 (K9) (9649 S), anti-Ac-Histone H3 (K18) (9675 S) antibodies from Cell Signaling Technology; anti-KAT3B/p300 (3G230), anti-Histone H3 (ab1791) antibodies from Abcam; anti-GAPDH (G8795), anti-β-actin (A5441), anti-FLAG (F3165), rabbit IgG (I5006) and mouse IgG (I5381) from Sigma‐Aldrich. Polyclonal antibodies against Smad4 pY195, pY301, or pY322 were generated by GL Biochem (Shanghai) Ltd. TGF-β (TGFB1-100) was purchased from StemRD, SB431542 (S4317) from Sigma‐Aldrich and Gleevec/imatinib (CDS022173) from Sigma‐Aldrich, respectively.

Wang L., Gu S., Chen F., Yu Y., Cao J., Li X., Gao C., Chen Y., Yuan S., Liu X., Qin J., Zhao B., Xu P., Liang T., Tong H., Lin X, & Feng X.H. (2023). Imatinib blocks tyrosine phosphorylation of Smad4 and restores TGF-β growth-suppressive signaling in BCR-ABL1-positive leukemia. Signal Transduction and Targeted Therapy, 8, 120.

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Transforming Growth Factor-β Signaling

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Recombinant human TGFB1 was obtained from R&D Systems as a Chinese hamster ovary cell-derived protein of more than 97% purity based on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Lyophilized TGFB1 was reconstituted in a solution of 4 mM HCl with 0.1% BSA as a carrier protein. The TGF-β type I receptor inhibitor SB431542 (S4317) was purchased from Sigma-Aldrich. Monoclonal mouse anti-α-tubulin antibody (sc-23948; diluted 1:2000) was obtained from Santa Cruz Biotechnology. The polyclonal rabbit anti-LOX (ab31238) antibody (diluted 1:1000) was obtained from Abcam. The horseradish peroxidase-conjugated goat anti-rabbit and goat anti-mouse immunoglobulin G antibodies were obtained from Bio-Rad Laboratories.

Fang Y., Chang H.M., Cheng J.C., Klausen C., Leung P.C, & Yang X. (2016). Transforming growth factor-β1 increases lysyl oxidase expression by downregulating MIR29A in human granulosa lutein cells. Reproduction (Cambridge, England), 152(3).

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