Fig. 4

Overexpression of SARS-CoV-2 genes induces cell death of hPSCs-CMs. A The scheme showing CM differentiation from hESCs in 2D monolayer condition, followed by TMT-MS assessments. Wild type CMs serve as control. B–D Signaling pathway enrichment analysis of the upregulated proteins induced by Nsp6^OE, Nsp8 ^OE and M ^OE in hESC-CMs. E–G GO analysis of the upregulated proteins induced by Nsp6^OE, Nsp8^OE and M^OE in hESC-CMs. H The scheme of CM differentiation from hPSCs in 3D embryoid body (EB) condition, followed with apoptotic assessments. I–J Flow cytometry analysis of TUNEL⁺ cells in the CM (CTNT⁺) population of hiPSCs-derived EBs. All bars are shown as mean ± SD. (n = 4). A two-tailed unpaired t test was used to calculate p values: *p < 0.05 (vs. Control). K Representative immunostaining images of TUNEL⁺ cells in hiPSCs-derived EBs containing CTNT⁺ CMs. Scale bar, 100 µm. L Statistical data analysis of immunostaining results from (K). All bars are shown as mean ± SD. (n = 3). A two-tailed unpaired t test was used to calculate p values: *p < 0.05 (vs. Control). M–N Western blotting detection of CASP3, cleaved CASP3, CASP9 and cleaved CASP9 protein expressions in hESC-CMs (M) and hiPSC-CMs (N).