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. 2016 Jul 27;12(10):1738–1758. doi: 10.1080/15548627.2016.1196318

Figure 7.

Figure 7.

Type I IFN production is required for the activation of the extrinsic apoptosis pathway. (A) PK-15 and 3D4/2 cells were transfected and infected as described in the legend to Fig. 2A and C, which were considered as the control groups. For inhibition of IFNA and IFNB1 induction, cells were cultured with bufexamac (10 μM) after 1 h of virus absorption, or cotransfected with shRNA targeting IFNB1 prior to CSFV infection. At 48 h after infection, the mRNA levels of TNFSF10, TNFRSF10A, FASLG, and FAS were measured by qRT-PCR. The data represent the mean ± SD of 3 independent experiments. One-way ANOVA test; test of homogeneity of variances, P > 0.05, LSD (L) was used for correction of post-hoc test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; #, P > 0.05. (B) PK-15 and 3D4/2 cells were treated as described in (A). After 48 h, the activity of CASP3, CASP8, and CASP9 in cultured cells was detected by the CASP activity assay as described in Materials and Methods. The data represent the mean ± SD of 3 independent experiments. One-way ANOVA test; test of homogeneity of variances, P > 0.05, LSD (L) was used for correction of post-hoc test. *, P < 0.05; **, P < 0.01; #, P > 0.05. (C) PK-15 and 3D4/2 cells were treated as described in (A). After 48 h, cell samples were analyzed by western blot with antibodies against PARP and ACTB (loading control). The relative expression ratios of the targeted proteins were analyzed by densitometric scanning. The data represent the mean ± SD of 3 independent experiments. One-way ANOVA test; test of homogeneity of variances, P > 0.05, LSD (L) was used for correction of post-hoc test. ***, P < 0.001. (D) PK-15 and 3D4/2 cells were treated as described in (A). After 48 h, IFNA and IFNB1 production and the inhibition efficiency of type I IFN induction were detected by the ELISA assay. The data represent the mean ± SD of 3 independent experiments. One-way ANOVA test; test of homogeneity of variances, P > 0.05, LSD (L) was used for correction of post-hoc test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; #, P > 0.05.