Fig. 1: Isolation of PDGFR endocytic vesicles utilizing magnetic nanoparticles.
From: Endocytic vesicles act as vehicles for glucose uptake in response to growth factor stimulation
a Schematic showing strategy for endocytic vesicle isolation. b Fractions magnetically isolated from post-nuclear supernatants of PDGF-BB-biotin-conjugated nanoparticle-treated (PDGF-particle) or unconjugated PDGF-BB plus nanoparticle-treated (control) Swiss 3T3 fibroblasts were analyzed by SDS-PAGE and silver staining. Post-nuclear supernatant (input), flowthrough, and wash fractions were from PDGF-BB-biotin-conjugated nanoparticle-treated cells. A representative image from one of 5 independent experiments is shown. c Venn diagram shows numbers of proteins identified in the control and endocytic vesicle fractions by LC-MS/MS, as detailed in Supplementary Data 1. Data are from a single experiment. d Gene ontology (GO) enrichment analyses of proteins uniquely identified in the endocytic vesicle fraction. Annotations related to endocytosis or vesicle transport are in red. One-sided P values were calculated using Fisher’s Exact test. e Fractions magnetically isolated from post-nuclear supernatants of PDGF-BB-biotin-conjugated nanoparticle-treated (PDGF-particle) or unconjugated PDGF-BB plus nanoparticle-treated (control) Swiss 3T3 fibroblasts were subjected to immunoblotting with the indicated antibodies. Representative images are shown from one of 2 independent experiments. f Heatmap representing numbers of peptide spectrum matches (#PSMs) of glycolysis-related proteins obtained by LC-MS/MS. A schematic of glycolysis is shown on the right. g Fractions magnetically isolated from post-nuclear supernatants of PDGF-BB-biotin-conjugated nanoparticle-treated (PDGF-particle) or unconjugated PDGF-BB plus nanoparticle-treated (control) Swiss 3T3 fibroblasts were subjected to immunoblotting with indicated antibodies. Representative images are shown from one of 2 independent experiments. Source data are provided as a Source Data file.
