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. 2021 Jul 22;12(8):728. doi: 10.1038/s41419-021-04011-0

Fig. 5. Afatinib-triggered autophagy plays a protective role in HNSCC.

Fig. 5

A–D FaDu and HN6 cells were pretreated with 5 mM 3-MA for 2 h (A, B) or with 10 μM CQ for 30 min (C, D) followed by co-treatment with 2 μM afatinib for another 24 h. The level of LC3B and apoptosis-associated proteins caspase-3 and PARP was measured by western blot analysis (A, C), while flow cytometry analysis was carried out to evaluate apoptosis (B, D). E, F FaDu and HN6 cells were transfected with control and Atg5 siRNA. Forty-eight hours after transfection, cells were exposed to 2 μM afatinib for 24 h. Atg5, LC3B, and apoptosis-associated proteins caspase-3 and PARP were measured by western blot analysis (E), meanwhile flow cytometry analysis was carried out to evaluate apoptosis (F). All data are presented as mean ± SD from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001.