Anti actb antibody

Total EPC protein was extracted and quantified by RIPA Lysis Buffer (Beyotime Biotechnology, China) containing protease inhibitors (Roche) and BCA assay kit (I Thermo, USA) separately. Protein extracts were separated by SDS-PAGE, transferred to PVDF membranes (Roche, Indianapolis, IN, USA). The following antibodies were used: anti-Beclin1 antibody (1 : 1000; ImmunoWay, USA), anti-Atg5 antibody (1 : 1000; ImmunoWay, USA), rabbit anti-LC3 antibody (1 : 1000; Cell Signaling Technology, USA), and anti-ACTB antibody (1 : 1000; Cell Signaling Technology, USA). Proteins were visualized with HRP-conjugated anti-rabbit IgG or anti-mouse IgG (1 : 3000; Cell Signaling Technology, USA), followed by the use of the ECL chemiluminescence system (Thermo). And the level of protein was analyzed by using Image J.

Nuclear extracts (10 μg) were subjected to SDS–PAGE, transferred to a polyvinylidene fluoride membrane that was blocked with 5% skim milk and then incubated with an anti-c-Fos antibody (sc-52, Santa Cruz Biotechnology, Inc., TX, USA, 1:1000) or an anti-ACTB antibody (#4970, Cell Signaling Technology, MA, USA, 1:1000). Full blotting images corresponding to the immunoblottings shown in the main figures are provided as Supplementary Fig. 1.