Figure 7
LDAH and ATGL display antagonistic effects. (A) Flag-LDAH and/or ATGL were transfected to HEK293 cells as indicated. TAG was measured after 24 h of OA (360 μM) loading and normalized to protein. ANOVA analysis confirmed a significant effect of both LDAH and ATGL on intracellular TAG stores at p < 0.001. Pairwise comparisons are shown in the chart. *p < 0.05, **p < 0.005; N.S. = not significant. (B–E) LDAH-GFP was co-transfected with Flag or Flag-ATGL to HEK293 cells, and cells were treated with OA (360 μM) for 24 h. (B) Average size of LDs per cell; (C) total area of LDs per cell; (D) number of LDs per cell; and (E) size distribution of LDs. Fifteen to sixteen cells per group were used for the measurements. *p < 0.05, **p < 0.005. (F) Diagram of experimental approach (top). LDAH-GFP transfected cells were treated with OA (360 μM) for 24 h. Cells were then transfected with ATGL-cherry in the presence of triacsin C (1 μg/ml). Expression of LDAH-GFP alone (a) and ATGL-cherry alone (c), and co-expression of LDAH-GFP and ATGL-cherry (c) were visualized by confocal microscopy. White arrows: LDs in cells expressing only LDAH-GFP. Yellow arrows: LDs in cells expressing both LDAH-GFP and ATGL-cherry. (G) Quantification of LD area per cell in cells overexpressing both LDAH-GFP and Cherry, or both LDAH-GFP and ATGL-Cherry (n = 14 per group). **p < 0.005.
