Treated MB cells were washed twice with ice-cold PBS and whole cell extracts (WCE) were prepared by lysing cells in modified RIPA buffer and fractionated in 4–20% Tris-Glycine Gels (Bio-Rad, Hercules, CA). Western Blot analysis were performed by transferring proteins to PVDF membranes (Millipore, Bedford, Mass) as previously described [24 (link), 25 (link)]. Membranes were probed using the appropriate primary and secondary antibodies, diluted according to the manufacturers’ instructions. Signals were detected by enhanced chemiluninescence and using MyECL imager (ThermoScientific, MA, USA).
Primary antibodies (Abs) used for Western blots are as follows:
STAT3 (sc-483 and sc-482), MYC (9E10, sc-40), BCL2 (sc-7382), CCND1 (sc-753), β-Actin (sc-47778), E-Cadherin (sc-21791), N-Cadherin (sc-271386), Vimentin (sc-6260), CHOP (sc-7351), PIAS3 (sc-46682) and OCT4 (sc-5279) were purchased from Santa Cruz (Dallas, TX).
P-Tyr 705 STAT3 (# 9145), PARP (# 9532), GAPDH (# 5174), Vinculin (# 3901), Cyclophilin (# 2175) were purchased from Cell Signaling (Danvers, MA). Nestin Ab (ABD69) was purchased from Millipore (Billerica, MA).
Primary antibodies (Abs) used for Western blots are as follows:
STAT3 (sc-483 and sc-482), MYC (9E10, sc-40), BCL2 (sc-7382), CCND1 (sc-753), β-Actin (sc-47778), E-Cadherin (sc-21791), N-Cadherin (sc-271386), Vimentin (sc-6260), CHOP (sc-7351), PIAS3 (sc-46682) and OCT4 (sc-5279) were purchased from Santa Cruz (Dallas, TX).
P-Tyr 705 STAT3 (# 9145), PARP (# 9532), GAPDH (# 5174), Vinculin (# 3901), Cyclophilin (# 2175) were purchased from Cell Signaling (Danvers, MA). Nestin Ab (ABD69) was purchased from Millipore (Billerica, MA).
