Fig. 2. CYT387 enabled serial propagation of HRV-C in human airway organoids.

a A schematic graph outlines the experimental procedure for panels (b and c), created with Biorender.com. b In the first (P1) passage, after pre-treatment with CYT387 or DMSO overnight, airway organoids were inoculated with HRV-C3 (n = 2). Culture media were harvested from infected airway organoids at indicated h.p.i. to detect viral replication. In the second (P2) passage, airway organoids pretreated with CYT387 or DMSO were inoculated with 50 µl P1 medium collected from CYT387- or DMSO-treated organoids, respectively (n = 2). Culture media were harvested at indicated h.p.i. to detect viral replication. c From P3, airway organoids pretreated with CYT387 or DMSO were inoculated with medium collected from CYT387-treated organoids at 100 viral gene copy/cell (P3, n = 2; P4 and P5, n = 3). The culture media were harvested at 96 h.p.i. to detect viral replication. The viral loads in P1 and P2 media at 96 h.p.i. are incorporated. Data represent mean and SD of the indicated number (n) of biological replicates from a representative experiment. Statistical significance (P4 and P5) was determined using a two-tailed Student’s t-test. **P < 0.01. ns not significant. Source data are provided as a Source Data file for Fig. 2b, c. d At 24 h.p.i. of HRV-C3, airway organoids were fixed and immune-labeled with an α-VP3 (green) and α-ACCTUB (red). Nuclei and actin filaments were counterstained with DAPI (blue) and Phalloidin-647 (white), respectively. The experiment was independently performed three times with similar results. Scale bar, 10 µm.