Fig. 4

CHML interacts strongly with Rab14. a silver stained gel shows differential bands between control and CHML sample. White arrow indicates differential band, the upper one indicates Flag-CHML. M marker, vec vector control. b Tandem mass spectrum analysis of target band. c, d Interaction between CHML and Rab14 was demonstrated by immunoprecipitation. 293T cells were transfected with Flag-CHML (c) or Flag-Rab14 (d). Immunoprecipitation was performed at 36 h post transfection with Flag antibody. e, f Western blot results showing endogenous CHML interacted with Rab14 in PLC/PRF/5 (e) and YY-8103 (f) cells. Black arrow indicates light chain. g In vitro interaction between CHML and Rab14. Purified 6xHis CHML was incubated with indicated GST fusion proteins, GST pull-down assay was conducted. h Graphic shows the domains in CHML protein. i In vitro interaction assay showing the two GDI domains interacted with Rab14. The truncated domains of CHML were fused with GST, and incubated with Rab14, followed by GST pull-down and WB