Fig. 4: Delivery of CSpV1-dsRNAs triggers activation of type I IFN signaling in host cells.

a Upregulation of type I IFN and type I IFN-stimulated genes in IEC4.1 cells transfected with CSpV1-dsCA or CSpV1-dsRdRp for 24 h as revealed by RT-qPCR. b Slight activation of type I IFN signaling in IEC4.1 cells transfected with CSpV1-ssRdRp-F, CSpV1-ssRdRp-R, CSpV1-ssCA-F and CSpV1-ssCA-R, respectively, for 24 h. Data (in a and b) are from three biological replicates and presented as mean values ± SD. p values were determined by one-way ANOVA followed by Tukey’s HSD test. c Heatmap of gene expression profiles in IEC4.1 cells following C. parvum infection or cells transfected with CSpV1-dsCA or CSpV1-dsRdRp or in combination. Cells were infected for 24 h or transfected with CSpV1-dsRNAs for 24 h. The heatmap representing differentially expressed genes (log10 fold changes) for each biological replicate (3 RNA-seq replicates each group). d Venn Diagrams demonstrating genes differentially expressed (with adjusted p values < 0.05) in IEC4.1 cells following infection (24 h p.i.) or transfected with CSpV1-dsCA or CSpV1-dsRdRp or their combination (for 24 h). e Heatmap representing expression of type I IFN-stimulated genes in infected IEC4.1 cells (24 h p.i.) or cells transfection of CSpV1-dsCA or CSpV1-dsRdRp or their combination (24 h). Data (in c–e) are from three RNA-Seq biological replicates for each group. f Interference with CSpV1-RNA delivery through transfection of host cells with siRNA combinations (siPOOLs) designed to target CSpV1-dsRNAs. IEC4.1 cells were transfected with the siPOOLs for 24 h, followed by infection for 24 h. Levels of CSpV1-dsRdRp were measured by RT-qPCR. A non-specific scrambled siRNA (siControl) was used for control. g Infection burden in IEC4.1 cells transfected with the siPOOLs or siControl for 24 h, followed by infection for 24 h. Levels of cpHsp70 were measured by RT-qPCR. h Attenuation of C. parvum-triggered Ifnb1 transcription in cells treated with siPOOLs. IEC4.1 cells were transfected with the siPOOLs or siControl for 24 h, followed by infection for 24 h, and Ifnb1 levels were measured. Data (in f–h) are from three biological replicates and presented as mean values ± SD. p values were determined by two-way ANOVA followed by Tukey’s HSD test (in f and h). Source data are provided as a Source Data file.