A new role of anterograde motor Kif5b in facilitating large clathrin-coated vesicle mediated endocytosis via regulating clathrin uncoating

a Silver-staining analysis of immune-isolated mouse cortical extracts by using Kif5b antibody or control IgG. The lower and upper band (arrowheads) were verified as Kif5b and CHC by mass-spectrometry. b, c Immunoprecipitation of Kif5b in mouse cortical extracts. The precipitates were analyzed by indicated antibodies. d Representative electron micrographs of cortical CCVs stained with control IgG isotype or antibody against Kif5b, α-Adaptin or Kif17, followed by incubation of 2nd antibody conjugated to gold particles. Arrowheads indicate the gold particles representing Kif5b or α-Adaptin on CCVs. Scale bar？=？100？nm. e Percentage of gold-positive CCVs in different samples. 305, 500, 324, and 1105 CCVs from images of 35, 46, 36, and 67 randomly-selected fields were analyzed for samples treated with Kif5b antibody, Kif17 antibody, control IgG isotype or α-Adaptin antibody, respectively. Error bars indicate s.e.m. **P？<？0.01. f Diameter distribution of analyzed CCVs for different groups. Data of 933 CCVs from Kif5b antibody-treated samples, 277 Kif5b gold-positive CCVs, 385 α-Adaptin gold-positive CCVs and 2733 overall CCVs were used for plotting. IB immunoblot, IP immunoprecipitation, Cla Clathrin light chain A, Clb Clathrin light chain B, Dyn I Dynamin I, Syn6 Syntaxin 6, SynTM Synaptotagmi