Fig. 4

HOTAIR regulates the expression and the localization of RAB35. a Real-time PCR analysis of the mRNA expression of RAB5, RAB7, RAB11, RAB27A, RAB27B, and RAB35, which encode GTPases involved in the release of exosomes, in HOTAIR overexpressing HepG2 cells. b Western blotting analysis of RAB35 protein levels from the above cells. c Confocal co-localization analysis of CD63 (red) and RAB35 (green) in HepG2 cells transfected with pcDNA3.1or pcDNA3.1-HOTAIR. The rectangular box indicates the small punctate structures where CD63 and RAB35 were co-localized. d RIP assays were performed using RAB35 antibody or control IgG antibody in HepG2 cells, and then real-time PCR analysis was used to measure the enrichment degrees of HOTAIR coprecipitated RNA. e NTA analysis of exosome secretion in HepG2 cells co-transfected with pcDNA3.1-HOTAIR and si-Rab35. Data are reported as the mean ± standard error (SD) from three independent experiments, t-test *P-value < 0.05