Fig. 5. S10A synapsin inhibits basal intracellular retention of Glut4 in adipocytes. L1 adipocytes differentiated on culture dishes were infected as described in Fig. 3. Cells were treated ± insulin (100 nM, 30 minutes), placed on ice, then labeled with anti-HA (detected with PC7). After washing, cells were lifted from the plate with collagenase at 4°C, and analyzed by flow cytometry. MFR, mean fluorescence ratio (PC7/GFP). 5000 cells/sample. Mean values from uninfected cells were used to correct for autofluorescence and non-specific binding. MFRs were standardized to G4 basal samples. (A) Average MFR ± s.d. from three experiments. (B) Average foldstimulation (MFR Insulin/MFR basal) ± s.d. from three experiments. (C) Comparison of the effects of S10A synapsin and S10A dsRED, a fusion protein with the first 118-amino acids of S10A synapsin fused to dsRED. White, basal; black, insulin; grey, ratio. Statistical analysis (Bonferroni, Scheffe's and Tukey tests; Origin 7.5) was done after standardizing to either the basal or insulin stimulated controls (^***P<0.01 significant difference; ns, no significant difference, P>0.05).