Fig. 4.

Bat intestinal organoids reproduced bat susceptibility to coronaviruses. a–c At the indicated hours after bat intestinal organoids inoculated with CoV-HKU4 (a), EV-71 (b) and human intestinal organoids inoculated with EV-71 (c), culture media were harvested and applied to viral load detection by RT-qPCR and viral titration by TCID50 assay. Data represent mean and s.d. in a representative experiment, n = 3. Two-tailed unpaired Student’s t test. d EV-71 infected and mock-infected human intestinal organoids were fixed and immunostained to identify viral protein VP1 positive (green) cells. Nuclei and actin filaments were counterstained with DAPI (blue) and Phalloidin-647 (purple), respectively. Scale bar, 20 µm. e At the indicated hours after human intestinal organoids inoculated with CoV-HKU4, culture media were harvested and applied to viral load detection and viral titration. Data represent mean and s.d. in a representative experiment, n = 3. Two-tailed unpaired Student’s t test. f Bat and human intestinal organoids infected with CoV-HKU4 or mock-infected were fixed and immunostained to identify CoV-HKU4 NP positive (green) cells. Scale bar, 20 µm. g, h At the indicated hours after human and bat intestinal organoids inoculated with WT SARS-CoV-2 and the Omicron variant, culture media were harvested and applied to viral load detection and viral titration. Data represent mean and s.d. in a representative experiment, n = 3. Ordinary one-way ANOVA with Tukey’s multiple comparison test. i SARS-CoV-2 infected and mock-infected bat and human intestinal organoids were fixed and immunolabeled to identify SARS-CoV-2 NP positive (green) cells. Scale bar, 20 µm