Fig 2: OAS-RNase L and other dsRNA pathways act together to induce the proinflammatory response(A) Schematic of RNA-seq experiments (n = 3–5 experiments).(B) Venn diagram showing the overlap of DE genes in poly(I:C)-treated WT (magenta), 2-5A-treated WT (yellow), and poly(I:C)-treated RNASEL-KO cells (green, abbreviated KO).(C) Correlation between differential expression changes in 2-5A- (RNase L alone) vs. poly(I:C)-treated cells. Blue and red dots represent JNK/p38 MAPK activation-related or IFN-stimulated genes, respectively. Black dot represents GADD34.(D) Comparison of JNK/p38 and IFN pathways under 2-5A (RNase L alone) vs. poly(I:C) activation. p < 0.001, indicated by three asterisks and calculated by using Student’s t test.(E) Effects of activating the OAS-RNase L pathway on phosphorylation of JNK (n = 5), p38 (n = 7), ZAKα (n = 3), eIF2α (n = 3), and IRF3 (n = 4) assayed by western blotting.(F) Quantification of blots in (E). Activation levels were normalized to that of poly(I:C)-treated WT cells. Bars represent standard error.(G) Model for RNase L acting synergistically with the other dsRNA pathways to enhance the antiviral effect for JNK and p38 via ZAKα. See also Figure S3 and Table S1.

Fig 3: Induction of apoptosis in HeLa cells by kuwanon C. (A) Apoptosis and necrosis staining was used to detect the apoptosis and necrosis level of HeLa cells treated with 0 μM (control), 30 μM, 60 μM of kuwanon C, albanin A, norartocarpetin, and paclitaxel for 24 h. (Red arrows indicate apoptotic cells, and white arrows indicate necrotic cells). Scale bar = 200 μm. (B) The impact of kuwanon C on the microstructure of HeLa cells. HeLa cells were treated with 0 μM, 60 μM, and 80 μM kuwanon C and examined using transmission electron microscopy. Scale bar = 10 μm. (C) Flow cytometry was performed to detect the apoptosis level of HeLa cells after treatment with different concentrations (control, 25 μM, 50 μM) of kuwanon C, albanin A (50 μM), and norartocarpetin (50 μM) for 24 h. (D,E) The expression levels of apoptosis-related genes (GADD45A, Casp3, Casp4, ERN1, PPP1R15A, PMAIP1, ATF3, and IFIT2) were quantified using (RT-qPCR). (F) Western blotting detection of protein levels of apoptosis-related proteins GADD34, NOXA. Each bar represents the mean ± SD of three independent experiments. Statistical significance is indicated by * p < 0.05, ** p < 0.01, *** p < 0.001 and ns: not significant.