Fig. 8

Effects of the mitochondrial KATP channel inhibitor 5-HD on the cell surface expression of GLUT1. HUVECs were cultured under normoxia or hypoxia with or without 5-HD (100 μM). a Cell surface expression of GLUT1 determined by flow cytometric analysis. The cells were labeled with a human anti-GLUT1 Alexa Fluor 700-conjugated antibody to detect cell surface GLUT1. Left panel: The histograms represent cell counts (Y-axis, linear scale) versus fluorescence intensity (X-axis, log scale). Right panel: Fluorescence mean intensity obtained from the histograms was quantified and shown in bar graphs (right panels). Data are the mean ± SEM from 4 independent experiments. *P < 0.01 versus normoxia control (Mann–Whitney test). b The cellular distribution of GLUT1 was determined by immunofluorescence staining as described in the Materials and Methods. HUVECs were stained with an anti-GLUT1 antibody (green). Left panel: Images were obtained with a confocal fluorescence microscope as described in the “Materials and methods”. Representative images are shown from 4 independent experiments with similar results. Scale bars, 25 μm. Right panel: The number of cells in which GLUT1 was observed at the cell membrane was counted in approximately 100 cells from 4 independent experiments. Data are the mean ± SEM from 4 independent experiments. *P < 0.01 versus normoxia control (Mann–Whitney test)