Immunofluorescence staining
Ba/F3 cells after 24 h treatment (10ug/mL JMT101,100 nmol/L EGFR TKIs or 100 nmol/L EGFR TKIs plus 10 ug/mL JMT101) were seeded on poly-L-lysine coated slides. The slides were fixed in 4% formaldehyde for 15 min, permeabilized with Triton X-100 (Beyotime) for 10 min, and incubated with primary antibody overnight at 4 °C. The primary antibodies used in the study were rabbit anti-EGFR (1:100; Cell Signaling Technology, #4267 S, Rabbit monoclonal [D38B1], lot:24) at a dilution of 1:100. The slides were rinsed twice with PBS, followed by incubation with the appropriate Alexa Fluor 488-labeled Goat Anti-Rabbit IgG (Beyotime) for 1 h at room temperature. The cells were counterstained with Antifade Mounting Medium with DAPI (Beyotime), and the coverslips were mounted on slides.