Fig. 4: Kif5b regulates large CCV-dependent VSV cellular entry.

a Representative immunofluorescence staining images of internalized VSVs in shKif5b or shControl Neuro2a cells after 0 or 15 min VSV infection. White dashed lines delineate the cell surface defined by co-staining of F-actin with phalloidion. Green pseudo color was used for VSV signals. Scale bar = 10 μm. b Quantitative analysis of internalized VSV signals for shKif5b or shControl Neuro2a cells infected by VSV with different time slots. c Representative immunofluorescence staining images for LDLR (green) in shKif5b or shControl RNA treated Neuro2a cells counterstained with DAPI (blue). Scale bar = 20 μm. d Western blot analysis to compare LDLR expression level in shKif5b or shControl RNA treated Neuro2a cells. e left column: representative immunofluorescence staining images of VSVs (Green) bound to cells counterstained with DAPI (blue). Scale bar = 20 μm. Right column: quantification of binding of VSVs to shKif5b or shControl RNA treated Neuro2a cells by FACS flow cytometry. Red dashed line indicates similar fluorescence value among histograms of different experimental groups. Neuro2a cells bound with viruses were stained with normal IgG isotype as a negative control. f Co-immunoprecipitation of CHC with Kif5b from Neuro2a cells with 1 h pre-treatment with or without 100 µM cell-penetrating scramble or Kif5b^891-915 peptide at 37 °C. g Representative immunofluorescence staining images of internalized VSVs in Neuro2a cells with 15 min infection after 1 h pretreatment without peptide or with 100 µM scramble or Kif5b^891–915 peptide at 37 °C. Scale bar = 10 μm. h Quantitative analysis of internalized VSV signal in different Neuro2a cell groups indicated in (g). Error bars indicate s.e.m. n > 30. ****P < 0.0001; ** P < 0.01; *P < 0.05. ns, not significant (P > 0.05)