Protein Expression Analysis of Cultured Cells

Cell lysis buffer (#R0010, Solarbio, China) was used to collapse the cells on the ice, phenylmethanesulfonylfluoride (PMSF, #P0100, Solarbio) was added in one hundredth of the volume of lysis buffer to restrain the degradation of protein. The protein concentration of lysates was quantified using the Enhanced BCA Protein Assay Kit (#P0010S, Beyotime, China). The western blot assay was strictly following the procedures of a previous publication^{21 (link)}. The primary antibodies included ALKBH5 Monoclonal antibody (#67811-1-Ig, Proteintech, China), VEGFA Monoclonal antibody (#66828-1-Ig, Proteintech, China), VAV1 Polyclonal antibody (#16364-1-AP, Proteintech, China), FGFR1 Monoclonal antibody (#60325-1-Ig, Proteintech, China), and GAPDH Polyclonal antibody (#10494-1-AP, Proteintech, China). The horseradish peroxidase (HRP)-conjugated affinipure goat anti-rabbit IgG (1:5000, SA00001-2, Proteintech, China) and HRP-conjugated affinipure goat anti-mouse IgG (1:5000, SA00001-1, Proteintech, China) served as corresponding secondary antibodies after incubation with primary antibody. The value of protein was calculated using the ImageJ software.