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

Fig. 1. Afatinib induces autophagy in HNSCC cells.

Fig. 1

A Western blot analysis of Beclin-1 and LC3B expression in FaDu, HN6, and CAL-27 cells after treatment with 0, 1, 2, and 4 μM afatinib for 24 h. The intensity of Beclin-1 and LC3B was normalized to β-actin, and the fold induction (treated/untreated control) was calculated (*p < 0.05, **p < 0.01, ***p < 0.001 vs. untreated control). B Western blot analysis of Beclin-1 and LC3B expression in FaDu, HN6, and CAL-27 cells after treatment with 2 μM afatinib for 0, 4, 8, 12, and 24 h. C, D FaDu-EGFP-LC3B and HN6-EGFP-LC3B cell lines were treated with 0, 1, 2, and 4 μM afatinib for 24 h (C) or treated with 2 μM afatinib for 0, 4, 8, 12, and 24 h (D). Then, cells were fixed, imaged, and EGFP-LC3B fluorescent spots were quantified. Scale bar, 25 µm. All data are presented as the mean ± SD from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001.