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. 2021 Jun 14;11:662444. doi: 10.3389/fonc.2021.662444

Figure 4.

Figure 4

DsRNA is necessary for siRNAs targeting mouse-specific lncRNA AA388235 to induce human tumor cell pyroptosis/apoptosis. (A, B) Microscopic images of HCT116 and DLD1 cells transfected as indicated. dsRNA, double-stranded siRNA; guide, single-stranded-anti-sense of siRNA; passenger, single-stranded sense of siRNA; guide + passenger, single-stranded-anti-sense of siRNA and single-stranded-sense of siRNA; dsDNA, double-stranded DNA which has the same sequence as siRNA. Red arrowheads indicated the pyroptotic cells, and yellow arrowheads indicated the apoptosis cells. Scale bar, 100 μm. (C, D) Immunoblotting assay of HCT116 and DLD1 cells transfected as indicated. g + p, guide + passenger, single-stranded-anti-sense of siRNA and single-stranded-sense of siRNA. GSDME-FL, full-length of GSDME; GSDME-N, the N-terminal cleavage of GSDME; Cleaved-CAS3, the cleavage of caspase-3 p19/p17; cleaved-PARP, the cleavage of PARP. (E, F) Microscopic images of HCT116 and DLD1 cells transfected with shRNA plasmids as indicated. Red arrowheads indicated the pyroptotic cells, and yellow arrowheads indicated the apoptosis cells. Scale bar,100μm. (G, H) Immunoblotting assay of HCT116 and DLD1 cells transfected with shRNA plasmids as indicated. sh-2, shAA388235-2; sh-6, shAA388235-6; GSDME-FL, full-length of GSDME; GSDME-N, the N-terminal cleavage of GSDME; Cleaved-CAS3, the cleavage of caspase-3 p19/p17; cleaved-PARP, the cleavage of PARP.