Fig. 2

SPRY2 KO increases glucose uptake and lipid droplet accumulation in HepG2 cells. a HepG2 mock, SPRY2 KO and SPRY2 OE cells were treated with 100 μg/mL 2-NBDG for 30 min. Cells were washed in PBS, treated with 3.3 μM Hoechst 33342 in PBS and imaged on the GFP and DAPI channels of the EVOS fluorescence microscope. The mean fluorescence per cell, representing glucose uptake, was calculated using CellProfiler and expressed relative to mock. Bars depict mean + SD from 3 independent experiments, with 2 to 4 wells per condition, 9–16 images per well. *P < 0.05 by one-way ANOVA with Bonferroni’s post-hoc test comparing KO with mock. b Cells were treated with Bodipy 493/503 (0.5 μg/mL) to label intracellular lipids and imaged using fluorescence microscopy. CellProfiler was used to calculate the mean GFP fluorescence per cell, representing lipid droplet accumulation. Bars depict mean + SD from 3 independent experiments, relative to mock, with 2 to 4 wells per condition, 9–16 images per well. **P < 0.01, ***P < 0.001 by one-way ANOVA with Bonferroni’s post-hoc test; ‘ns’ denotes not significant