Fig. 2: OSBP translocates from Golgi to PM via interacting with ORP4L.

a Electron micrographs of ORP4L KI and wild-type T-cells, HepG2 and 293 T cells. Scale bar, 100 nm. Black triangles present in dividual data points for n = 20 cells from one mouse or cell line. The central mark indicates the median, the bottom and top edges of the box indicate the interquartile range, and the whiskers represent the maximum and minimum point. One-way ANOVA test with a confidence interval of 95% was used to compute statistics. b Co-immunoprecipitation analysis of ORP4L binding to OSBP in ORP4L KI T-cells with or without anti-CD3 stimulation. c The localization of OSBP in ORP4L KI T-cells. Scale bar, 10 μm. d The localization of OSBP in ORP4L KI and wild-type T-cells with or without anti-CD3 stimulation. Scale bar, 10 μm. e Western blot analysis of OSBP protein levels in PM and Golgi of ORP4L KI or wild-type T-cells. f A schematic representation describing the BiFC technique. g Interactions between ORP4L/pVn-C1 and OSBP/pVc-C1 in ORP4L KI T-cells as determined by BiFC. Scar bar, 10 μm. h The localization of OSBP in ORP4L KI T-cells upon ORP4L knockdown. Scar bar, 10 μm. i Western blot analysis of OSBP protein levels in PM and Golgi of ORP4L KI T-cells upon ORP4L knockdown. j The localization of overexpressed wild-type OSBP or OSBP (△ORP4L) in ORP4L KI T-cells. Scar bar, 10 μm. k Western blot analysis of OSBP protein levels in PM and Golgi of ORP4L KI T-cells with or without VAPA knockdown. l Western blot analysis of overexpressed wild-type OSBP or OSBP(△FFAT) protein levels in the PM and Golgi of ORP4L KI T-cells. Confocal microscopy and blot images are representative of n = 3 biological replicates with similar results. The same experiments were repeated twice in T-cells from two mice. In (c, d, h and j), the ration of OSBP fluorescence density from n = 10 cells from one mouse between PM and Golgi are shown below. Source data are provided as a Source Data file.