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. 2020 Nov;8(22):1502. doi: 10.21037/atm-20-7145

Figure 4.

Figure 4

Overexpression of BNIP3 in D407 cells alleviated the impairments of hypoxia. (A) Decreased expression of BNIP3 was investigated with quantitative RT-PCR (top) and western blotting (bottom). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as an internal control. (B) Quantitative RT-PCR (top) and western blotting (bottom) results to verify the overexpression of BNIP3 in D407 cells after infection with BNIP3 or lentivirus vector. GAPDH was used as an internal control. (C) BNIP3 overexpression promoted the cell growth in hypoxic conditions, which was inhibited by the administration of 50 µM 3-MA. (D) Hypoxia induced apoptosis in D407 cells, which was inhibited by BNIP3 overexpression. (E) Overexpression of BNIP3 recovered the inhibited autophagy by hypoxia to some extent. The immunofluorescence of LC3 (green) in D407 cells with or without BNIP3 overexpression was measured with fluorescence microscopy. The nuclei were stained with 4',6-diamidino-2-phenylindole (DAPI) in blue. Scale bar: 50 μm. (F) Overexpression of BNIP3 alleviated the impairments of hypoxia. Asterisks indicate significant difference (***, P<0.001).