Antibodies and western blotting
MUTANT
For western blotting, EGFR mutant Ba/F3 cells were incubated with compound (10 ug/mL JMT101, 100 nmol/L EGFR TKIs or 100 nmol/L EGFR TKIs plus 10 ug/mL JMT101) for 6 hours and evaluating levels of pEGFR in cellular lysates using Western blotting for pEGFR, EGFR, pERK1/2, ERK1/2, pAKT, and AKT with antibodies from Cell Signaling Technology. Primary antibodies included rabbit anti-pEGFR (1:1000; Cell Signaling Technology, #3777 S, Rabbit monoclonal [D7A5], lot:16), rabbit anti-EGFR (1:1000; Cell Signaling Technology, #4267 S, Rabbit monoclonal [D38B1], lot:24), rabbit anti-pAKT (1:1000; Cell Signaling Technology, #9271 S, lot:15), rabbit anti-AKT(1:1000; Cell Signaling Technology, #4691 S, Rabbit monoclonal [C67E7], lot:28), rabbit anti-pERK (1:2000; Cell Signaling Technology, #4370 S, Rabbit monoclonal [D13.14.4E], lot:24), rabbit anti-ERK (1:1000; Cell Signaling Technology, #4695 S, Rabbit monoclonal [137F5], lot:28)]. Blots were probed with rabbit antibodies against GAPDH (1:10000; Proteintech, #10494-1-AP) as a loading control. Source data are provided in the Source Data file.