Sb431542

Manufactured by ReproCELL

SB431542 is an inhibitor of the Transforming Growth Factor-beta (TGF-β) signaling pathway. It selectively inhibits the activin receptor-like kinase (ALK) family of receptors, particularly ALK4, ALK5, and ALK7, which are involved in the TGF-β signaling cascade. This compound can be used in cell culture and research applications to study the role of the TGF-β pathway in various biological processes.

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Lab products found in correlation

Ldn193189, by Merck Group (1 mentions) Glutamax supplement, by Thermo Fisher Scientific (1 mentions) Mem neaa, by Thermo Fisher Scientific (1 mentions) Knockout serum replacement (ksr), by Thermo Fisher Scientific (1 mentions) β mercaptoethanol, by Merck Group (1 mentions) Dmem f12, by Caisson (1 mentions) Basic fibroblast growth factor (bfgf), by STEMCELL (1 mentions) U bottomed plates, by Thermo Fisher Scientific (1 mentions) Ldn 193189, by ReproCELL (1 mentions) Matrigel, by Corning (1 mentions) Accutase, by STEMCELL (1 mentions) Bone morphogenetic protein 4 (bmp4), by R&D Systems (1 mentions) Vegf a, by Thermo Fisher Scientific (1 mentions) Rock inhibitor y 27632, by Selleck Chemicals (1 mentions) Geltrex, by Thermo Fisher Scientific (1 mentions) Chir99021, by Thermo Fisher Scientific (1 mentions) Vitronectin, by STEMCELL (1 mentions) Activin a, by R&D Systems (1 mentions)

3 protocols using sb431542

Feeder-Free Pluripotent Stem Cell Culture

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DMEM/F-12 (Caisson Labs, Cat No. DFL15)

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Knockout serum replacement (Thermo Fisher, Cat No. 10828-028) 15% (v/v)

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MEM-NEAA (Thermo Fisher, Cat No. 11140050) 1% (v/v)

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Glutamax supplement (Thermo Fisher, Cat No. 35050061) 1% (v/v)

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β-Mercaptoethanol (Sigma, Cat No. M7522) 100 μM

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LDN-193189 (Sigma, Cat No. SML0559) 100 nM

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SB431542 (Reprocell, Cat No. 04001005) 10 μM

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XAV939 (Stemgent, Cat No. 040046) 2 μM

Ressler A.K., Sampaio G.L., Dugger S.A., Sapir T., Krizay D., Boland M.J., Reiner O, & Goldstein D.B. (2022). Evidence of shared transcriptomic dysregulation of HNRNPU-related disorder between human organoids and embryonic mice. iScience, 26(1), 105797.

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Differentiation of ESCs into Organoids

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Induction followed the protocol of Koehler and Hashino (2014) (link), but with modifications (Figure S7). In brief, ESCs were dissociated in Accutase (STEMCELL Technologies) and resuspended in differentiation medium (DMLK; Table S6). On D0, 1,500 cells in 100 μL of DMLK per well were plated in low binding 96-well U-bottomed plates (Thermo Fisher). On D1, half of the medium was exchanged with fresh DMLK containing Matrigel (Corning; 2% final concentration). Bone morphogenetic protein 4 (PromoKine) and SB-431542 (Reprocell) were added on D3. Later, basic fibroblast growth factor (STEMCELL Technologies) and LDN-193189 (Reprocell) were added (Figure S7). On D8, aggregates were washed twice in PBS before being transferred to new 96-well U-bottomed plates in 100 μL of N2 medium (Table S6) containing 1% Matrigel and 3 μM CHIR99021 (DeJonge et al., 2016 (link)). After 48 h, aggregates were transferred to 24-well low binding plates in fresh N2 medium until D20. On D20, aggregates were cultured in organoid medium (Table S6) with constant shaking. Half of the medium was changed every other day during the long-term culture period.

Tang P.C., Alex A.L., Nie J., Lee J., Roth A.A., Booth K.T., Koehler K.R., Hashino E, & Nelson R.F. (2019). Defective Tmprss3-Associated Hair Cell Degeneration in Inner Ear Organoids. Stem Cell Reports, 13(1), 147-162.

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Endothelial Cell Differentiation Protocol

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Endothelial cells were differentiated as previously described^{76 (link)} with minor changes. hESCs were plated on Geltrex (ThermoFisher (Gibco), A1413302) coated plates in TeSR-E8TM media (StemCell Technologies, 05990) with 1 μM Rock inhibitor (Y-27632, Selleckchem, S1049). Two days later (d0), cells were collected and reseeded onto non-treated culture dishes coated with 1 μg cm⁻¹ Vitronectin (StemCell Technologies, 07180) at a density of 100,000 cells per cm² in TeSR-E8 medium supplemented with 5 ng ml⁻¹ BMP4 (R&D, 314-BP-050/CF), 25 ng ml⁻¹ Activin A (R&D, 338-AC-050/CF) and 1 μM CHIR99021 (PeproTech, 2520691) for 2 d. At d2, the media were switched to TeSRTM-E7TM media (StemCell Technologies, 05914) supplemented with 50 ng ml⁻¹ VEGF-A (Peprotech, 100-20), 50 ng ml⁻¹ BMP4 (R&D, 314-BP-050/CF) and 5 μM SB431542 (ReproCell, 04-0010-05) for another 4–5 d. Final differentiated cells were stained with antibodies against CD31 (Pe-Cy7; Biolegend,303118) and CD144 (PE; Biolegend, 138009).

Harding J., Vintersten-Nagy K., Yang H., Tang J.K., Shutova M., Jong E.D., Lee J.H., Massumi M., Oussenko T., Izadifar Z., Zhang P., Rogers I.M., Wheeler M.B., Lye S.J., Sung H.K., Li C., Izadifar M, & Nagy A. (2023). Immune-privileged tissues formed from immunologically cloaked mouse embryonic stem cells survive long term in allogeneic hosts. Nature Biomedical Engineering, 8(4), 427-442.

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