Fig. 5: TRIM39 decreases the ubiquitination of Rab7 at lysine 191 residue.

a Vector or FLAG-TRIM39 was co-transfected with HA-ubiquitin into HCT116 cells for 48 h. Cell lysates were extracted and immunoprecipitated using an anti-Rab7 antibody. Then the precipitates were examined with anti-HA. b shNC and TRIM39 stably knocked down HCT116 cells were transfected with HA-ubiquitin for 48 h. Cell lysates were extracted and immunoprecipitated using an anti-Rab7 antibody. Then the precipitates were examined with anti-HA. c Myc-Rab7^K191R was co-transfected with HA-ubiquitin into shNC or TRIM39 stably knocked down HCT116 cells for 48 h. Cell lysates were immunoprecipitated with anti-Myc antibody and immunoblotted with anti-HA and anti-Myc antibody. d GST-RILP^RBD fusion protein immobilized on glutathione-Sepharose beads were incubated with Myc-Rab7^WT or Myc-Rab7^K191R overexpressed HCT116 cell lysates at 4 °C for 4 h. Rab7 and GST were detected in the washed beads by western blot. e shNC and TRIM39 stably knocked down HCT116 cells were transfected with Myc-Rab7^K191R for 48 h. Cell lysates were immunoprecipitated with anti-Myc antibody and immunoblotted with anti-GDI2 and anti-Myc antibody. f HCT116 cells were transfected with Myc-Rab7^WT or Myc-Rab7^K191R for 48 h. Cell lysates were immunoprecipitated with anti-Myc antibody and immunoblotted with anti-MON1A and anti-Myc antibody. g shNC or TRIM39 stably knocked down HCT116 cell lysates were extracted and immunoprecipitated with anti-Rab7 antibody. MON1A was detected by western blot.