Purified macrophage-lineage cells were cultured in serum-free DMEM for 72-96h for SR-A1 inhibition with 10 μM rhein (Sigma) and/or for collecting culture media/cell lysates for ELISA/immunoblotting, or in DMEM/5% FBS for 48-120h for 50% HEK293T-CM (Mock or STC1-CM) and/or 75 μg/ml fucoidan (Santa Cruz) treatment to induce macrophage differentiation. Mouse primary lung fibroblasts enriched by serial passaging of lung tissues (Figure S3 ) were subjected to modified 3T3 culture, in which 3 × 105 cells were re-plated every 4-6 days in DMEM/10% FBS, or co-cultured for 72h with IMCs in serum-free medium for TGFβ1 ELISA or in DMEM/F12 containing 10%FBS ± 1 μM SB431542 (Cambridge Bioscience) for BrdU uptake assay. HEK293T cells were maintained in DMEM/10%FBS.
Sb431542
Manufactured by Cambridge Bioscience
SB431542 is a selective inhibitor of the transforming growth factor-beta (TGF-β) type I receptor ALK5, ALK4, and ALK7. It is a widely used tool compound in cell biology research.
Automatically generated - may contain errors
Lab products found in correlation
Fucoidan, by Santa Cruz Biotechnology (1 mentions)
Rhein, by Merck Group (1 mentions)
A83 01, by Merck Group (1 mentions)
Matrigel, by BD (1 mentions)
L ascorbic acid, by Bio-Techne (1 mentions)
Valproic acid, by Merck Group (1 mentions)
Mtesr1, by STEMCELL (1 mentions)
Chir99021, by Bio-Techne (1 mentions)
Y 27632, by STEMCELL (1 mentions)
2 protocols using sb431542
1
Macrophage Differentiation and Co-culture
2
Directed Trophoblast Stem Cell Reprogramming
Check if the same lab product or an alternative is used in the 5 most similar protocols
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H9 hESCs cells were routinely cultured in mTeSR1 (Stem Cell Technologies; Cat. No. 85850) and growth factor reduced Matrigel (BD Biosciences; Cat. No. 356321). For hTSC reprogramming experiments, H9 cells were plated in mTeSR1 media on Matrigel coated cell culture dishes. Media was changed for hTSC media on the first day of reprogramming. hTSC media was prepared as described previously (Okae et al., 2018) (Chir99021, BioTechne, Cat. No. 4423/10; EGF, Tebu Bio Ltd, Cat. No. 167AF-100-15-a; ITS-X supplement, Fisher Sci, Cat.
No. 10524233; L-ascorbic acid, Tocris, Cat. No. 4055/50; A83-01, Sigma-Aldrich Cat. No. SML0788/5MG; SB431542, Cambridge Bioscience, Cat. No. SM33-2; Valproic acid, Sigma-Aldrich Cat. No. V-006-1ML; Y27632, Stem Cell Technologies, Cat. No. 72302). Media was changed every second day. For transgene induction, doxycycline was added daily at a concentration of 1 μg/ml over 20 days for transgene induction. hTSCs were passaged by dissociation with TrypLE Express (Fisher Thermo Scientific, Cat. No. 12604013) for 15 min at 37 C and passaged onto collagen IV-coated (5 µg/ml; Corning, Cat. No. 354233) 10 cm plates and maintained in hTSC media.
No. 10524233; L-ascorbic acid, Tocris, Cat. No. 4055/50; A83-01, Sigma-Aldrich Cat. No. SML0788/5MG; SB431542, Cambridge Bioscience, Cat. No. SM33-2; Valproic acid, Sigma-Aldrich Cat. No. V-006-1ML; Y27632, Stem Cell Technologies, Cat. No. 72302). Media was changed every second day. For transgene induction, doxycycline was added daily at a concentration of 1 μg/ml over 20 days for transgene induction. hTSCs were passaged by dissociation with TrypLE Express (Fisher Thermo Scientific, Cat. No. 12604013) for 15 min at 37 C and passaged onto collagen IV-coated (5 µg/ml; Corning, Cat. No. 354233) 10 cm plates and maintained in hTSC media.
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