Fig. 4

Effect of PP1α and PP1γ silencing on Glucose uptake and GLUT4 translocation in insulin sensitive and insulin resistant condition: Proliferated N2a cells were transfected with non-specific (scrambled) and PP1α and PP1γ specific siRNA. Post transfection cells were differentiated in the absence (MF; insulin sensitive) or chronic presence of 100 nM insulin (MFI; insulin resistant) for 3 days. A and B For glucose uptake assay, transfected N2a cells were serum-starved for 2 h and treated with 100 nM insulin for 30 min. Treated cells were lysed and uptake of 2-NBDG was then measured. Bar represents relative change in the uptake of 2-NBDG. Data expressed are mean ± SE. ***p < 0.001 compared to Lane 1, ^###p < 0.001 compared to Lane 2, ^θθp < 0.01 compared to Lane 4 and ^δδp < 0.001 compared to Lane 6. Open bars: MF, filled bars: MFI, A.U. Arbitrary Units, SC Scrambled C For GLUT4 translocation, post transfection, MF MFI differentiated N2a cells were treated with or without 100 nM insulin for 30 min followed by fixation and permeabilization. Cells were subjected to immunofluorescence microscopy by using anti-goat Alexa 555 secondary antibody. Images were captured from different fields and a representative image of three images is presented. Scale Bar: 100 μm. White arrow: GLUT4 redistribution around the nucleus; Yellow arrows: GLUT4 redistribution/translocation on the plasma membrane