Fig. 4: Virological features of BA.2.86/JN.1 and EG.5.1/HK.3 in human nasal epithelial cells.

a Replication of BA.2, BA.2.86, JN.1, EG.5.1 and HK.3 in Calu3 cells (n = 6). Cell lysates were quantified for viral subgenomic E gene (sgE) copies. The n number represents biological repeats. b Replication of BA.2, BA.2.86, JN.1, EG.5.1 and HK.3 in hNECs (n = 3). Cell lysates were quantified for viral sgE copies. The n number represents biological repeats. c Pseudovirus entry in hNECs. hNECs were transduced with BA.2.86- (n = 7), JN.1- (n = 7), EG.5.1- (n = 4), and HK.3-spike (n = 4) pseudoviruses. Pseudovirus entries were quantified by measuring the luciferase signal. The n number represents biological repeats. d Preference of protease usage by BA.2.86 and JN.1 in hNECs (n = 3). The supernatant was quantified for viral RdRp gene level with qRT-PCR. The n number represents biological repeats. e Spike cleavage of BA.2.86, JN.1, EG.5.1, and HK.3 in hNECs. A representative image of the spike was shown with β-actin added as a sample processing control. Spike and β-actin were run on different gels and detected on different membranes. The cleavage ratio of spike proteins was quantified by ImageJ. f Quantification of spike cleavage as performed in Fig. 4e (n = 4). The n number represents biological repeats. g Infectivity of BA.2.86, JN.1, EG.5.1, and HK.3 in hNECs. Infected hNECs were fixed at 48 hpi for the visualization of ciliated cell marker beta-tubulin (red) and SARS-CoV-2 nucleocapsid protein (green). The DAPI channel was included in the merged images. Scale bar, 20 μm. h Immunofluorescence signal intensity of SARS-CoV-2 N BA.2.86 (n = 5), JN.1 (n = 9), EG.5.1 (n = 7), and HK.3 (n = 5) in Fig. 4g was quantified with ImageJ. The n number represents biological repeats. Scale bar = 20 µm. Data represent mean ± SEM. Statistical significance in (a–c and f, h) was determined with one-way ANOVA with post hoc Tukey’s multiple comparisons test. Statistical significance in (d) was determined with two-way ANOVA with Sidak’s multiple comparison tests. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. NS is not statistically significant. Source data are provided as a Source Data file.