Fig. 3: Infection and proliferation of N3-R. heilongjiangensis.
a N3-R. heilongjiangensis was purified from azide-labeled Vero-81 cells, and it emitted green fluorescence signals after the reaction between DBCO-AF 488 and azide moieties through click chemistry. N3-R. heilongjiangensis was stained with mouse polyclonal antibodies against R. heilongjiangensis (red), then the fluorescence signals of DBCO-AF 488 (green) and R. heilongjiangensis (red) were observed by a fluorescence microscope. WT-R. heilongjiangensis served as the control. Scale bar = 6.4 μm. b Infection and proliferation of Vero-81 cells infected with N3-R. heilongjiangensis at 72, 96, and 120 h post inoculation (hpi). N3-R. heilongjiangensis was purified from azide-labeled Vero-81 cells, then infected Vero-81 cells on slides in 24-well plates. N3-R. heilongjiangensis emitted green fluorescence signals after the reaction between DBCO-AF 488 and azide moieties through click chemistry. Simultaneously, N3-R. heilongjiangensis was stained with mouse polyclonal antibodies against R. heilongjiangensis (red), and the nuclei were stained with Hoechst 33342 (blue) at 72, 96, 120 hpi, then DBCO-AF 488 (green) and R. heilongjiangensis (red) signals were observed with a fluorescence microscope. WT-R. heilongjiangensis served as the control. Scale bar = 20 μm. c Comparison of growth between WT-R. heilongjiangensis and N3-R. heilongjiangensis in Vero-81 cells. Growth curves of WT-R. heilongjiangensis and N3-R. heilongjiangensis in Vero-81 cells were analyzed using quantitative PCR. n = 3 biologically independent samples. Error bars represent geometric means ± 95%CI. The blue dots and lines indicate the copy number changes of WT-R. heilongjiangensis. The green dots and lines indicate the copy number changes of N3-R. heilongjiangensis. Mann–Whitney U test was used for comparison among groups. N3-R. heilongjiangensis compared with WT-R. heilongjiangensis at the same hpi. P (2 h) = 0.002, P (12 h) = 0.002, P (24 h) = 0.026, P (48 h) = 0.002, P (72 h) = 1, P (96 h) = 0.053, P (120 h) = 0.454. **P < 0.01; *P < 0.05; NS, P > 0.05, no significance.
