Western Blot Analysis of MAPK Signaling

Cells after nsPEF stimulation were collected at 0.5 h and lysed by RIPA lysis buffer (R0020, Solarbio). The western blotting was performed according to the manufacturer’s protocol [13 (link)]. Rabbit polyclonal antibodies against Phospho-P38 MAPK (4511, Cell Signaling), P38 MAPK (8690, Cell Signaling), ERK1/2 MAPK (4695, Cell Signaling), Phospho-ERK1/2 MAPK (4370P, Cell Signaling), JNK MAPK (9252, Cell Signaling), Phospho-JNK MAPK (4668, Cell Signaling), CREB (4820, Cell Signaling), Phospho-CREB (9198, Cell Signaling), STAT3 (4904, Cell Signaling), Phospho-STAT3 (9145, Cell Signaling), β-catenin (sc-7199, Santa Cruz Biotechnology), and β-actin (13E5, Cell Signaling) were utilized to detect the targeted proteins, followed by incubation with secondary HRP-linked antibody of anti-rabbit IgG (Cell Signaling). The complex of the antigen and the antibody was detected with TANON 1600 Gel Imaging System, and the expression level of protein is analyzed with Tanon Gis. Statistical significance was marked with different letters (P < 0.05).