Fig. 2: TGF-β-induced palmitoylation promotes HK1 translocation to the plasma membrane.
For the experiments described in this figure, LX-2 cells were treated with TGF-β (2 ng ml−1) for 36 h as indicated, and cell lysates and lEVs were prepared for western blotting unless specifically indicated otherwise. a, Detection of HK1 palmitoylation in the presence of 2-BP (100 μM) (top) or PalmB (50 μM) (bottom) with or without TGF-β treatment. b,c, Effect of 2-BP and PalmB on HK1 translocation, analyzed by fractionation assays (b) and confocal microscopy (c; n = 3 independent experiments). Scale bar, 10 μm. d, Effect of 2-BP and PalmB on HK1 secretion. e–h, Comparison of HK1 palmitoylation (e), localization (f, g; n = 3 independent experiments) and secretion (h) in HK1-expressing and HK1 6CS-expressing LX-2 cells. Scale bar, 10 μm. i, Detection of HK1 palmitoylation (right) and secretion (left) in ABHD17B knockdown cells. j, Effect of TGF-β on ABHD17B mRNA and protein expression levels (n = 3 independent experiments). Flotillin-2 was used as a loading control for EVs. Tubulin was used as a protein loading control. Statistical data are presented as the mean ± s.e.m. Statistical analyses were determined by two-tailed Student’s t-test (c, right; j) and one-way ANOVA, followed by Tukey’s post hoc test (c, left; g). All western blots were repeated three times and one of them is shown.
