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

Fig. 6.

Fig. 6

SARS-CoV-2 spike induces inflammation and apoptosis through enhanced autophagy. (a-c) The protein expression levels of IL-6, IL-8, and TNF-α in SARS-CoV-2 spike pseudovirions-infected HEK293T, HEK293T-hACE2, Vero E6, 16HBE, and HMEC-1 cells pretreated with DMSO or 3-MA (10 mM) were quantified using ELISA. (d) Semiquantitative RT-PCR analysis of IL-6, IL-8, and TNF-α expression in SARS-CoV-2 spike pseudovirions-infected HEK293T, HEK293T-hACE2, Vero E6, 16HBE, and HMEC-1 cells pretreated with DMSO or 3-MA (10 mM). (e, f) Western blot analysis of Bcl-2 and Bax in SARS-CoV-2 spike pseudovirions-treated, 3-MA-treated, and SARS-CoV-2 spike pseudovirions and 3-MA (10 mM)-treated HEK293T, HEK293T-hACE2, Vero E6, 16HBE, and HMEC-1 cells. GAPDH was used as the loading control. Bcl-2/GAPDH and Bax/GAPDH densitometric ratios were recorded. (g) Apoptotic cells were determined using Annexin V-FITC/7-AAD double staining in SARS-CoV-2 spike pseudovirions-treated, 3-MA (10 mM)-treated, and SARS-CoV-2 spike pseudovirions and 3-MA (10 mM)-treated HEK293T, HEK293T-hACE2, Vero E6, 16HBE, and HMEC-1 cells. (h) Schematic illustrating the mechanism of SARS-CoV-2 spike to promote inflammation and apoptosis in infected cells. SCV-2-S, SARS-CoV-2 spike pseudovirions. All results were collected in three independent experiments. Data represent the mean ± SD (*P < 0.05; **P < 0.01; ***P < 0.001).