Sma ab5694

Proteins were separated by 10% SDS-PAGE (Solarbio, S1052) and then transferred onto a polyvinylidene difluoride (PVDF) membrane. The membranes were incubated with relevant antibodies in WB primary antibody diluent overnight at 4°C, including TGF-β1 (ab179695), α-SMA (ab5694), smad2 (ab63576), and smad3 (ab40854), which were purchased from Abcam (Cambridge, MA); anti-Smad7 (sc-11392) which was purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA), and rabbit anti-GAPDH (AB-P-R-001) which was obtained from GOOD HERE Technology. Then, membranes were incubated with horseradish peroxide-conjugated secondary antibodies for 1 hour at room temperature after washing. Proteins were visualized by an enhanced chemiluminescence system (Thermo Fisher Scientific Inc., Waltham, MA, USA). Band intensities were analyzed by Quantity One 1-D Analysis software (Bio-Rad Laboratories Inc., Hercules, CA, USA), and values were normalized against the value of GAPDH.

UCB-MSCs were acquired from Kang Stem Biotech (Seoul, Korea). Fetal bovine serum (FBS) and antibiotics were purchased from Hyclone (Logan, UT, USA) and Gibco (Grand Island, NY, USA), respectively. 4-P-PDOT (#SML1189), cholesterol (#C4951), DIDS (#D3514), melatonin (#M5250), and Ver155008 (#SML0271) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Mithramycin A (#1489) was purchased from Tocris (Minneapolis, MN, USA). Antibodies against ABCA1 (#ab18180), α-SMA (#ab5694), and Sp1 (#ab227383) were purchased from Abcam (Cambridge, England) and against HSF1 (#H00003297-A01) were purchased from Abnova (Taipei, Taiwan). Antibodies against NRF1 (#8052S) and cleaved-caspase 3 (#9661) were purchased from Cell Signaling Technology (Beverly, MA, USA), and those against BiP (#MA5-27686) and phospho-Sp1 (Thr453, #PA5-38333) were purchased from Invitrogen (Carlsbad, CA, USA). Antibodies against MT1 (#NBP1-71113) were purchased from Novus Biologicals (Littleton, CO, USA). Antibodies against β-actin (#sc-47778), caspase 9 (#sc-8355), lamin A/C (#sc-20681), and MT2 (#sc-28453) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). All primers for mRNA and miRNA were purchased from Cosmogenetech (Seoul, Korea). Small interfering RNAs (siRNAs) were purchased from Bioneer (Deajeon, Korea).