Rabbit anti beta tubulin

Western blot analysis was performed to determine the protein expression as previously described (33 (link)). The total proteins were extracted with radioimmunoprecipitation assay (RIPA) lysis buffer containing protease and phosphatase inhibitor cocktail (Beyotime), and quantified using BCA Kit (Beyotime), followed by separation on SDS-PAGE and transferring to polyvinylidene difluoride membranes (Millipore, Billerica, MA, USA), which were then reacted with different primary antibodies at 4 °C overnight. The primary antibodies included mouse anti-myosine heavy chain (MHC) antibody (R&D Systems, Minneapolis, MN, USA), rabbit anti-MuRF1, rabbit anti-MAFbx (Fbx32), rabbit anti-LC3B, rabbit anti-PINK1, rabbit anti-BNIP3, rabbit anti-ATG7, rabbit anti-Beclin 1, rabbit anti-beta tubulin (Abcam, Cambridge, UK), mouse anti-pJak1 (Tyr1034/1035)/Jak2 (Tyr1007/1008), rabbit anti-pSTAT3 (Tyr705), and rabbit anti-Stat3 (Cell Signaling Technology, Danvers, MA, USA). Then, the membranes were incubated with horse radish peroxidase (HRP)-conjugated secondary immunoglobin G (IgG) antibodies for 2 h. The target proteins were detected using enhanced chemiluminescence (Thermo Fisher Scientific, USA), and the band intensity was quantified and normalized against loading control.