Fig. 4: Autophagy was induced by prolonged hypoxia in HMEC-1 cells. | Cell Death & Disease

Fig. 4: Autophagy was induced by prolonged hypoxia in HMEC-1 cells.

From: Ischemia induces autophagy of endothelial cells and stimulates angiogenic effects in a hindlimb ischemia mouse model

Fig. 4

a HMEC-1 cells were exposed to 1% O2 for 2 h, 24 h, and 48 h. Western blotting of whole cell lysates was performed using the indicated antibodies. b The relative expression of p62 was quantified and plotted. **P < 0.01 and ***P < 0.001 versus normoxic conditions. c HMEC-1 cells were exposed to hypoxia for 2 and 24 h, and LC3 puncta (green) and p62 (red) expression was determined by immunofluorescent staining. Nuclei were counterstained with Hoechst 33342. Scale bar = 40 μm. d The white dotted square in c was magnified to examine the LC3 puncta. Scale bar = 20 μm. e The relative p62 fluorescence intensity of c was quantified using ImageJ software. Quantitative values were normalized to 100% normoxic HMEC-1 cells. ***p < 0.001 versus normoxia. f HMEC-1 cells were treated with hypoxia and subjected to CYTO-ID (green) staining. Cell nuclei were stained with Hoechst 33342. Scale bar = 40 μm. g HMEC-1 cells were transfected with the mCherry-EGFP-LC3B expression vector, and the cells were observed under a fluorescence microscope after hypoxia treatment. Scale bar = 40 μm.

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