P stat3 sc 8059

The adenovirus-infected cells were seeded in 6-well plates and treated with WT sericin hydrogel or 3.44 × 10⁻³μg/mL PDGF-BB sericin hydrogel. Cells were harvested at 24 h, 48 h, 9 days or 11 days after treatment. Cell lysates were separated by 10% SDS-PAGE (Beyotime) and transferred to PVDF (Polyvinylidene fluoride) membrane (Bio-Rad, CA, USA). The membrane was pre-blocked with 5% bovine serum albumin buffer for 60 min, probed with the primary antibody (1:1,000), washed three times with Tris-buffered saline containing 0.1% Tween-20 and then incubated with horseradish peroxidase-conjugated secondary antibodies. Primary antibodies for GAPDH (sc-32233), osteopontin (OPN) (sc-10593), Runx2 (sc-12488), osteocalcin (OCN) (sc-365797), peroxisome proliferators activated receptor-γ (PPARγ) (sc-7273), CCAAT/enhancer binding protein α (C/EBP α) (SC-61), Smad 1/5/9 (AF0614), p-Smad 1/5/9 (AF8313), Stat3 (60199-1) and p-Stat3 (SC-8059) were purchased from Santa Cruz Biotechnology (China). Assays were done in triplicate.

GGA was purchased from Sigma‐Aldrich (6809‐52‐5, St. Louis, MO, USA). Furamidine dihydrochloride (DB75, HY‐110137) was obtained from MedChemExpress (Monmouth Junction, NJ, USA). Antibodies recognizing Hsp70/Hsc70 (sc‐65521), PRMT1 (sc‐166963), CHIP (sc‐66803), Ubiquitin (sc‐8017), STAT3 (sc‐8019), p‐STAT3(sc‐8059) and Hsp 70 siRNA (sc‐29352) were purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, CA, USA). Anti‐FAS antibody (ab82419) and anti‐mono and dimethyl‐arginine antibody (ab412) were obtained from Abcam (Cambridge, UK). Antibodies against PRMT1 (#2449S), cleaved caspase‐3 (#9664), caspase‐3 (#9662), cleaved caspase‐8 (#9748), caspase‐8 (#4790), cleaved caspase‐9 (#9505), caspase‐9 (#9508), Bax (#14796), Bcl‐2 (#15071), Cyt c (#11940), p53 (#9282), Lamin B (#13435) and GAPDH (#2118) were purchased from Cell Signaling Technology, Inc (Beverly, MA, USA).