Fig. 6. V_m facilitates cell surface retention of nutrient transporters by configuring plasma membrane PIP₂ dynamics.

a Schematic of cell membrane protein extraction for quantitative proteomics analysis. b, c GO (b) and KEGG pathway (c) enrichment analysis of differentially expressed membrane proteins in peritoneal macrophages. d Confocal microscope analysis of membrane GLUT1 and 4F2hc internalization. (left, n = 10, 11, 15 respectively; right, n = 12, 14, 18 respectively). e, f Western blots of membrane GLUT1 expression in peritoneal macrophages treated with 500 ng/ml LPS in the presence or absence of 25 μM ML133 or 6 μg/ml nystatin for 2 h. g In vitro glucose uptake assays in mouse peritoneal macrophages. (n = 3, mean ± SD). h Western blots of pro-IL-1β expression in mouse peritoneal macrophages. i Western blots of pro-IL-1β expression in BMDMs overexpressing vector or the Grp1^DD mutant treated with or without 500 ng/ml LPS in the presence or absence of 25 μM ML133 for 6 h. j–l Confocal microscope analysis of membrane PI(4, 5)P2 clustering in mouse iBMDMs. (left, the 2D and 2.5D images of PI(4, 5)P2 staining; right, membrane fluorescence of PI(4, 5)P2) (j). Statistics of membrane clustering of PI(4, 5)P2 showing better clustering of PIP2 (k) and 3D reconstruction of membrane PIP2 (l). (n = 13, 19, 23 respectively for the left panel; n = 13, 15, 14 respectively for the right panel; mean ± SD). m In vitro glucose uptake assays and IL-1β transcription analysis. (n = 3, mean ± SD). Two-tailed unpaired Student’s t-test. The qPCR, western blot, and FACS data are representative of two or three independent experiments. Source data are provided as a Source Data file.