Raf 1

The expression of proteins in Huh7 and Huh7‐R cells was detected by performing western blot (WB). Total protein from Huh7 cells was extracted applying Total Protein Extraction Kit (Cat: BC3710; Solarbio) as the protocol described. BCA Protein Assay Kit (Cat: PC0020; Solarbio) was used to assess the protein concentration. Protein samples were electrophoresed on 10% SDS/PAGE gels. Then, the separated proteins were transferred onto polyvinylidene fluoride membranes (Cat: ISEQ00010; Merck Millipore, Billerica, MA, USA). Subsequently, the membranes were incubated with the primary antibodies, CXCR4 (1 : 1000, Cat: 11073‐2‐AP; Proteintech, Wuhan, China), Cav‐1 (1 : 1000, Cat: 16447‐1‐AP; Proteintech), p‐Cav‐1 (1 : 2000, Cat: ab75876; Abcam), c‐Met (1 : 1000, Cat: 25869‐1‐AP; Proteintech), p‐c‐Met (1 : 1000, Cat: ab68141; Abcam), Raf‐1 (1 : 1000, Cat: 26863‐1‐AP; Proteintech) and EGFR (1 : 10 000, Cat: 66455‐1‐Ig; Proteintech) at 4 °C for 12 h. After blocking with 5% skim milk, the membranes were incubated with goat anti‐rabbit or goat anti‐mouse horseradish peroxidase‐conjugated secondary antibody (1 : 5000, Cat: SA00001‐2/SA00001‐1; Proteintech). GAPDH antibody (1 : 5000, Cat: 10494‐1‐AP; Proteintech) was used as a reference protein for normalization. The data were analyzed by imagej software (National Institutes of Health, Bethesda, MD, USA).