Figure 8

Effects of vimentin on the internalization of NDV. HD11 cells in 6-well cell culture plates were pretreated with siRNA-VIM or siRNA-NC. The pretreated cells were infected with 1 MOI JS/7/05/Ch, JS/MukHN or Mukteswar at 4 ℃ and 37 ℃ for 1 h, successively. The mock-infected cells were assayed in parallel as a control. The infected cells were washed with ice-cold acidic PBS (pH = 1.5) to remove the attached but not yet internalized virions. The cells were then incubated with a mouse anti-NDV NP primary antibody and secondary antibody conjugated to FITC. The NDV-internalized cells were finally detected by flow cytometry, and flow cytometry scatter plots were generated using CytExpert 2.3 software. Positive cell populations were established based on granularity (SSC-A) and the FITC-A fluorescence channel. The ratio of the number of positive cells to the total number of cells was calculated and is displayed within the box. The statistical graphs were generated using GraphPad Prism 9.0 software. ****p < 0.0001; ns indicates no significance.