Fig. 4: Endocytic vesicles act as vehicles for glucose uptake.
From: Endocytic vesicles act as vehicles for glucose uptake in response to growth factor stimulation
a Serum-starved Swiss 3T3 fibroblasts were pre-treated with dynein inhibitors Ciliobrevin D (30 μM) or Dynarrestin (30 μM) for 20 min or left untreated and subjected to glucose uptake assay with or without PDGF-BB (50 ng/ml) in the presence of 2DG for 10 min. Graph shows 2DG6P normalized to total cellular proteins. Bars and error bars show means of 3 independent biological replicates (deep-color dots, biological replicates, n = 3) and ±SD. Values of biological replicates are means of technical replicates (light-color dots). P values were calculated using one-way ANOVA and post-hoc Tukey’s test. b Swiss 3T3 fibroblasts ectopically expressing mCherry-fused PDGFR (magenta) with or without Green Glifon4000 glucose sensor or EGFP (green) were serum-starved and stimulated with PDGF-BB (50 ng/ml) for 10 min. Higher magnification images of the boxed regions are shown. Representative data are shown from one of 5 independent live cell imaging experiments. Scale bar: 20 μm. c Serum-starved Swiss 3T3 fibroblasts were subjected to glucose uptake assay with PDGF-BB (50 ng/ml), EGF (50 ng/ml), bFGF (50 ng/ml), HGF (50 ng/ml), insulin (50 ng/ml) or unstimulated. Graph shows 2DG6P normalized to total cellular proteins. Bars and error bars show means of 3 independent biological replicates (deep-color dots, biological replicates, n = 3) and ±SD. Values of biological replicates are means of technical replicates (light-color dots). P values were calculated using one-way ANOVA and post-hoc Tukey’s test. d Post-nuclear supernatants and endocytic vesicle fraction with or without 1% Triton X-100 were subjected to glycolysis assay. Graph shows lactate production normalized to protein levels in each fraction. Representative data are shown from one of 2 independent experiments. Source data are provided as a Source Data file.
