Chemidoctm analysis system

After the DPCs were transfected for 36 h, the protein of DPCs were disposed using RIPA Lysis Buffer (Beyotime, Shanghai, China), protein concentrations were measured using Enhanced BCA Protein Assay Kit (Beyotime, Shanghai, China). The proteins were separated by SDS-PAGE, transferred to polyvinylidene difluoride (PVDF) membranes, and probed with 1:500 rabbit anti-BMP7 (Huabio, Hangzhou, China) and 1:2500 rabbit anti-GAPDH (Proteintech Group, Rosemont, IL, USA) antibodies, and then with 1:3000 goat anti-rabbit IgG HRG antibodies (ABclonal, Wuhan, China). Signals were detected with the enhanced chemiluminescence ECL Western Blot kit (Biosharp, Hefei, China). All experimental producers were performed per the manufacturer’s instructions. Protein was detected and analyzed by the ChemiDoc^TM Analysis System (Bio-Rad, Hercules, CA, USA).

After 48 h of transfection, the DPCs proteins were disposed of with RIPA lysis buffer (Beyotime, Shanghai, China), and concentrations were detected using the BCA method. The proteins were separated and then transferred to PVDF membranes, which were probed with 1:500 rabbit anti-EGR1 (Affinity, Melbourne, Australia), 1:2500 mouse anti-GAPDH (ABclonal, Wuhan, China), 1:1000 rabbit anti-PCNA (Abcam, Cambridge, UK), 1:1000 rabbit anti-CDK2 (Abcam, Cambridge, UK), 1:3000 goat anti-rabbit IgG HRG antibody (ABclon, Wuhan, China), and 1:3000 goat anti-mouse (ABclonal, Wuhan, China). The protein expressions were measured using the ECL Western Blot kit (BioSharp, Hefei, China), and analysed by the ChemiDoc^TM Analysis System (Bio-Rad, Hercules, CA, USA).