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. 2021 Aug 27;1867(12):166260. doi: 10.1016/j.bbadis.2021.166260

Fig. 2.

Fig. 2

ACE2-mediated SARS-CoV-2 spike pseudovirions infection induces autophagic responses. (a) Western blot analysis of ACE2 expression in HEK293T-hACE2 stable cells. GAPDH was used as the loading control. ACE2/GAPDH densitometric ratios were recorded. (b) Flow cytometric analysis of cell surface expression of ACE2 on HEK293T-hACE2 stable cells. Rabbit IgG was used as an isotype control. (c) Western blotting for autophagy markers LC3 and p62 after treatment with SARS-CoV-2 spike pseudovirions for 24 h. LC3-II/GAPDH and p62/GAPDH densitometric ratios were recorded. (d) HEK293T, HEK293T-hACE2, Vero E6, 16HBE and HEMC-1 were transfected with mCherry-GFP-LC3 followed by SARS-CoV-2 spike pseudovirions treatment for 24 h. GFP-LC3 and mCherry-LC3 puncta were visualized using confocal microscopy. Hoechst 33342 was used to stain the nuclei of cells (blue) (Scale bar, 10 μm). (e, f) Statistical bar charts of autophagosome and autolysosome, as observed in SARS-CoV-2-S pseudovirions-infected HEK293T, HEK293T-hACE2, Vero E6, 16HBE, and HMEC-1 cells. SCV-2-S, SARS-CoV-2 spike pseudovirions. All results were determined from three independent experiments. Data represent mean ± SD (*P < 0.05; **P < 0.01; ***P < 0.001).