Fig. 2

Lonidamine enhances M1 virus infection by inhibiting the antiviral immune response in a manner independent of HK2. a The HCT 116 cell line was treated with siRNAs targeting HK2 for 48 h. M1 virus (MOI = 1 pfu/cell) was added for another 24 h, and the infection rate of M1 virus (GFP percentage) was then determined by flow cytometry. n = 3. Statistical analysis was performed by one-way ANOVA with Dunnett’s test for pairwise comparisons. The knockdown efficiency of the siRNAs (48 h) targeting HK2 in the HCT 116 cell line was determined by western blotting. The relative expression levels of HK-2 compared to those in the control group were calculated and are labeled. GSEA of the IFN-α response gene set (b) and IFN-β gene set (c) in HCT 116 cells after treatment with M1 (MOI = 1 pfu/cell) or M1 (MOI = 1 pfu/cell) plus lonidamine (50 μM) for 24 h. The NES, p and FDR values are indicated in each box. d Heatmap of the top 20 IRGs inhibited by lonidamine plus M1 compared with M1 (LM/M) in HCT 116 cells. e The HCT 116 cell line was treated with control, M1 (MOI = 1 pfu/cell), lonidamine (50 μM) or M1 (MOI = 1 pfu/cell) plus lonidamine (50 μM) for 24 h, and the relative expression levels of the top 10 IRGs (ADM11, SECTM1, MYC, PLEKHA4, FKBP5, UCP2, ETV6, SAT1, IFIT1, and C10orf10) noted in (d) were determined by qPCR; n = 3. Statistical analysis was performed by one-way ANOVA with Dunnett’s test for pairwise comparisons. f Summary of the top 10 IRGs inhibited by the combination of lonidamine plus M1 compared with M1 alone (LM/M) in (e, f). g–i The HCT 116 cell line was treated with siRNAs targeting ADM11, SECTM1, and MYC for 48 h. M1 virus (MOI = 1 pfu/cell) was added for another 24 h, and the M1 virus infection rate (GFP percentage) was determined by flow cytometry. n = 3. Statistical analysis was performed by one-way ANOVA with Dunnett’s test for pairwise comparisons. The error bars indicate the mean ± SD values from three independent experiments with three technical replicates. ns nonsignificant; *p < 0.05, **p < 0.01, ***p < 0.001