Fig. 5: Recruitment of FBXW8 to the Golgi via interaction with PtdIns4P.
From: Glucose controls lipolysis through Golgi PtdIns4P-mediated regulation of ATGL
a, Representative photomicrographs of the immunofluorescence of CUL7, FBXW8 and ectopically expressed FBXW8–FLAG in HEK293-AAV cells stained by the indicated antibodies. Scale bar, 5 μm. Repeated independently three times with similar results. b,c, Immunoblot of CUL7 and FBXW8 in the WCE, Golgi and vesicle fractions extracted from HEK293T cells after glucose withdrawal or UCB9608 treatment (n = 6 independent experiments per group). Asterisk indicates a non-specific band. d, Schematic illustration of the domains of human FBXW8. e, Blotting analysis of PIP strips incubated with recombinant proteins FBXW8–FLAG and FBXW8 mutant–FLAG. Three times repeated independently with similar results. f, Immunoblot of recombinant protein FBXW8–FLAG and FBXW8 mutant–FLAG after pull-down by PtdIns4P agarose beads. Three times repeated independently with similar results. g,h, Immunoblot of FBXW8–FLAG and FBXW8 mutant–FLAG in the WCE, Golgi and vesicle fractions extracted from HEK293T cells after glucose withdrawal (n = 6 independent experiments per group). i, Schematic diagram illustrating the working model of the mechanism by which glucose availability is coupled to lipolysis via Golgi PtdIns4P. Results are shown as mean ± s.e.m. and analysed using two-tailed paired t-test (b, c, g and h). Source numerical data and unprocessed blots are available in .
