Anti p raf1

We performed Western blotting according to the protocol described in a previous study [51 (link)]. The following antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA) for Western blotting: anti-cleaved Caspase3 (Cat#9661), anti-cleaved Caspase9 (Cat#9509), anti-cleaved PARP-1 (Cat#9541), anti-JAK1 (Cat# 3344), anti-p-JAK1 (Cat#74129), anti-STAT3 (Cat#9139), anti-p-STAT3 (Cat#9145), anti-p-PI3K (Cat#4228), anti-p-RAF1 (Cat#9427), anti-p-ERK (Cat#4370), anti-AKT (Cat#4691), and anti-p-AKT (Cat#4060). The anti-14-3-3-θ (Cat# ab183075) antibody was purchased from Abcam (Cambridge, UK). The anti-GAPDH (Cat#AC002) antibody was purchased from ABclonal Technology (Wuhan, China). HRP-conjugated anti-rabbit IgG (Cat#AS014) and HRP-conjugated anti-mouse IgG (Cat#AS003) (ABclonal Technology, Wuhan, China) were used as secondary antibodies, and enhanced chemiluminescence reagent (Vazyme, Nanjing, China) was used for detection of signals after exposure of the membrane in an image analyzer (Bio-Tanon, Shanghai, China).

Ovarian cancer cell lines (SK-OV-3, OVCAR3 and A2780) were purchased from Type Culture Collection of Chinese Academy of Sciences and maintained according to their recommendations. The ERK inhibitor PD98059 was obtained from Sigma. Antibodies were obtained from the following companies: anti-RASAL2 antibody from Abcam; anti-β-actin antibody from Sigma; anti-E-cadherin, anti-vimentin, anti-claudin-1, anti-N-cadherin, anti-ZO-1, anti-Snail, anti-Slug, anti-Raf-1, anti-p-Raf-1, anti-MEK1/2, anti-p-MEK1/2, anti-ERK1/2 and anti-p-ERK1/2 antibodies from Cell Signaling.