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. 2023 Apr 1;299(5):104668. doi: 10.1016/j.jbc.2023.104668

Figure 6.

Figure 6

SARS-CoV-2 infection triggers GSDMD-mediated pyroptosis, which was reduced by inhibition of Hsp90. THP-1-ACE2 macrophages were infected with SARS-CoV-2 GFP/ΔN virus at an MOI = 10 for 4 h in the presence 17-DMAG. A, cells were lysed and levels of caspase-1, GSDMD, cleaved GSDMD, Hsp70, SARS-CoV-2 N, and β-actin were determined by Western blot. The protein level of GSDMD and cleaved GSDMD were quantified by immunoblot scanning and normalized with respect to β-actin. B, qRT-PCR for quantification of TNF-α and IL-1β mRNA in THP-1-ACE2 macrophages. C, cells were stained with Calcein/PI and DAPI, or tested by LDH assay (D). E, ELISA was performed to determine IL-1β and TNF-α levels in cell supernatants; ∗p ≤ 0.05; ∗∗p ≤ 0.01; n.s.: Not significant. ACE2, angiotensin converting enzyme 2; Hsp90, heat shock protein 90; LDH, lactate dehydrogenase; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2.