Mouse anti human gapdh antibody

The western blotting analysis was used to detect protein KLF9 and β-actin expression. RIPA cell lysate containing protease inhibitor was used to lyse the cells for 30 min. After centrifugation at 12,000 g for 15 min at 4 °C, the supernatant was obtained. After electrophoresis, the protein sample was transferred to a PVDF membrane, and the 5% skim milk powder was sealed at room temperature for 1 h. Rabbit antihuman KLF9 antibody (Proteintech, Chicago, USA), PARP antibody (Proteintech, Chicago, USA) or mouse anti-human GAPDH antibody (Proteintech, Chicago, USA) were added separately and incubated overnight at 4 °C. After washing with TBST, membranes were incubated with secondary antibody at room temperature for 2 h. TBST was used to clean the membrane 3 times. Bands were detected using the ECL Kit and β-actin was used as a loading control.

The western blotting analysis was used to detect protein MUC15, PARP and GAPDH expression. The proteins were collected as previous described 20 (link). RIPA cell lysate containing protease inhibitor was used to lyse the cells for 30 min. After centrifugation at 12,000 g for 15min at 4 °C, the supernatant was obtained. After electrophoresis, the protein sample was transferred to a PVDF membrane, and the 5% skim milk powder was sealed at room temperature for 1 h. Rabbit antihuman MUC15 antibody (Abcam, Cambridge, MA), PARP antibody (Proteintech, Chicago, USA) or mouse anti-human GAPDH antibody (Proteintech, Chicago, USA) were added separately and incubated overnight at 4 °C. After washing with TBST, membranes were incubated with secondary antibody at room temperature for 2 h. TBST was used to clean the membrane 3 times. Bands were detected using the ECL Kit and b-Actin was used as a loading control.

HO8910 miR-4461 sponge or A2780 cells miR-4461 sponge and their control cells were planted into six-well plates for 48 h. Cells were immersed in RIPA cell lysate containing protease inhibitor for 30 min, and the supernatant was acquired after centrifugation. Total protein concentration was measured using the BCA kit (Pierce; Thermo Fisher Scientific, Inc.) according to the manufacturer's protocol. After electrophoresis, the protein sample was transferred to a PVDF membrane, blocking with the 5% skim milk. Rabbit antihuman PTEN antibody (Proteintech, Chicago, USA), PARP antibody (Proteintech, Chicago, USA), or mouse anti-human GAPDH antibody (Proteintech, Chicago, USA) were added separately and incubated overnight at 4°C. membranes were incubated with secondary antibody at room temperature for 2 h. Chemiluminescent signals were detected by the ECL Kit and b-Actin was used as internal control.