Figure 5.

Regulatory enzymes involved in pyruvate/acetyl-CoA production in adipocytes promote survival of early germline cysts. (A) Western blots showing expression of Hex-C and ATPCL::GFP in dissected fat bodies at 12 hr and 3 days on rich vs. poor diets. The expression levels on a poor diet were normalized to those on a rich diet for each time point. Total loaded proteins are shown with MemCode stain. (B) Western blotting analysis of Hex-C and ATPCL::GFP in fat body at 7 days of Lsp2^ts-mediated RNAi against Hex-C and ATPCL, respectively. Total proteins are used as a loading control (bottom). (C) Percentage of germaria containing cleaved Dcp1-positive cysts at 0, 7, or 14 days of Lsp2^ts-mediated RNAi against Hex-C, Pepck, ATPCL and AcCoAS. The number of germaria analyzed from three independent experiments is shown inside or to the right of bars. *P < 0.05, **P < 0.01, Chi-square test. (D–F) Germaria of females subjected to 14 days of Lsp2^ts-mediated luciferase (luc) control (D), Hex-C (E), or Pepck (F) RNAi knockdown. Hts (green), fusome; Lamin C (green), cap cell nuclear envelope; cleaved Dcp1 (red), dying cystoblasts/cysts; DAPI (blue), nuclei. Arrows indicate dying germ cells. Scale bar, 10 μm. (G) TAG contents (μg TAG/μg protein) in dissected fat bodies at 7 days of Lsp2^ts-mediated RNAi against luc control, Hex-C, Pepck, AcCoAS and ATPCL. **P < 0.01, Student’s t-test. (H–M) Adipocytes at 7 days of Lsp2^ts-mediated RNAi against luc control, Hex-C, Pepck, AcCoAS and ATPCL. DAPI (blue), nuclei; Phalloidin (green), actin; Nile Red (red), lipid droplets. Scale bar, 40 μm. AcCoAS, acetyl-CoA synthase; ATPCL, ATP citrate lyase; Hex-C, hexokinase-C; luc, luciferase; Pepck, phosphoenolpyruvate carboxykinase; RNAi, RNA interference; TAG, triacylglycerol.