Fig. 5.

PIP₂ add-back restores the GlcN-induced PIP₂ decrease and protects against GlcN-induced F-actin loss and insulin resistance. During the final 60 min of control or GlcN incubation, the medium was replaced with the same medium enriched with either histone H1 (A, panels 1, 2, 4, and 5; C and D, panels 1 and 2) or PIP₂/histone H1 (A, panels 3 and 6; C and D, panels 3). Representative immunofluorescent (IF) images of PM PIP₂ (A, panels 1–3) and WGA-stained PM (A, panels 4–6), and quantitation of PM PIP₂ (B) are shown. Phalloidin-stained F-actin and propidium iodide-stained nuclei in these cells (C), insulin-stimulated PM GLUT4 images (D), and GLUT4 quantitation (E) are shown. All microscope and camera settings were identical between groups and images and means ± se are from three to five independent experiments. *, P < 0.05 vs. control; #, P < 0.008 vs. 30′ Ins.).