Fig. 3.

Antiviral activity of gemcitabine against rotavirus SA11 in human intestinal organoids. (A) Human intestinal organoids (HIOs) were inoculated with SA11 rotavirus strain and treated with gemcitabine (GCT) for 48 h. The relative cellular rotavirus RNA and secreted viruses calculated as genomic copy number were analyzed by qRT-PCR (n = 6). Standard curve for calculation of genomic copy number is included in Supplementary Fig. S3. (B) The expression of viral structural protein VP4 was stained and quantified by Western blot assay (n = 4). (C) Optical microscopy images, confocal images and indirect fluorescence microscope images of HIOs from mock group and GCT treated groups. In confocal images, VP6 protein was stained in red, green signal represents Epcam, and nuclei were visualized by DAPI (blue). In fluorescence microscope images, HIOs were stained by PI (red) indicating dead cells, Hoechst (blue) for nuclear, and Calcein (green) as live cells. (D) Rate of deteriorated HIOs was calculated as dead organoids/total organoids (n = 6). Data represent means ± SEM; *P < 0.05; **, P < 0.01.