Ab225958

Briefly, NPC cells were collected and then lysed using lysis buffer for extraction of total proteins. Next, the proteins were separated on dodecyl sulfate, sodium salt-Polyacrylamide gel electrophoresis (SDS-PAGE, 10%), transferred to PVDF members (Millipore, IPVH00010), and incubated using primary antibodies (1:1000; Abcam, Cambridge, MA, USA) against ELF2 (ab225958) and β-actin (ab8227). Finally, the proteins were incubated with the second antibodies (Abcam, ab205718) and analyzed by the ECL plus Kit (Beyotime, Haimen, China, P0018S).

First, each group of cells was harvested, and then proteins were extracted using a lysis buffer. Next, we quantified the proteins with a BCA protein reagent kit (Thermo Scientific, USA). Equal amounts of protein extractions were isolated by SDS‐PAGE gels and transferred onto polyvinylidene fluoride (PVDF) membranes at 300 mA for 90 min and incubated overnight with primary antibodies at 4°C (1:1000; Abcam, Cambridge, MA, USA) against SMAD2 (ab225958) and β‐actin (ab8227). Last, we incubated proteins with the second antibodies (Abcam, ab205718) and analyzed proteins with the ECL plus kit (Beyotime, Haimen, China, P0018S). Images were gained with a BioSpectrum 600 Imaging System (UVP, CA, USA). This experiment was repeated three times and the average value was taken.