Fig. 4: Ggpps regulates hepatocyte-derived EV secretion.

a EV production by primary hepatocytes from WT and liver-specific Ggpps knockout mice (LKO) fed a normal chow diet or a HFD. (n = 6 mice per group). b CD63, TSG101 and Calnexin protein levels in parental cells and EVs from WT and LKO mice fed a normal chow diet or a HFD. c Membrane-associated Rab27A (lipid-soluble protein with geranylgeranylation in the detergent phase, down) and cytoplasm-associated Rab27A (water-soluble protein with no geranylgeranylation in the aqueous phase, up) were obtained by Triton X-114 extraction and analysed by immunoblotting. (n = 3 biologically independent samples). d Membrane-associated Rab5 (lipid-soluble protein with geranylgeranylation in the detergent phase, down) and cytoplasm-associated Rab5 (water-soluble protein with no geranylgeranylation in the aqueous phase, up) were obtained by Triton X-114 extraction and analysed by immunoblotting. (n = 3 biologically independent samples). e Fluorescence microscopy of primary hepatocytes co-infected with empty vector or Ggpps vector and WT or mutant Rab27A and stained with anti-CD63 antibodies and DAPI (nucleus, blue). (Scale bar, 5 μm). f EV production by primary hepatocytes from e. (n = 6 biologically independent samples). g TSG101 protein levels in the parental cells and EVs from e. Data from 8-week-old WT and LKO mice fed a normal chow diet or a HFD for 12 weeks are presented as the means ± SEM. Groups were analysed using an unpaired t test, *P < 0.05, **P < 0.01, ***P < 0.001, WT vs. LKO; ^###P < 0.001, Chow vs. HFD. Source data are provided as a Source Data file. See also Supplementary Fig. 5.