Fig. 2.

DOC2B enrichment preserves insulin-stimulated glucose uptake via GLUT4 translocation in L6-GLUT4-myc myotubes exposed to insulin resistance stimuli. (a, b) Expression of DOC2B and transduction efficiency for L6-GLUT4-myc myoblasts that were differentiated into myotubes and transduced with Ad-DOC2B or vector control adenoviral particles at multiple MOIs, as shown by (a) western blot analysis and (b) evaluation of GFP fluorescence (from viral packaging); scale bar, 400 μm. White arrows denoting DOC2B-transduced myotubes in the fluorescence image correspond to the same myotubes seen in the bright field image. (c) 2-DG uptake in myotubes incubated under control or InsRes conditions. The myotubes were transduced with Ad-GFP or Ad-DOC2B before induction of InsRes conditions. (d) Surface GLUT4 levels in L6-GLUT4-myc myoblasts transfected with DOC2B-GFP or control GFP plasmid DNA, incubated under control or InsRes conditions and measured in response to stimulatory insulin. For (c) and (d), the cells were stimulated with 100 nmol/l insulin to induce 2-DG uptake or GLUT4 translocation. The immunofluorescence intensity of cell-surface GLUT4 was normalised to the nucleic acid staining dye SYTO 60. n=3. *p<0.05, **p<0.01, ***p<0.001 as shown. Ctrl, control; Ins, insulin