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. Author manuscript; available in PMC: 2015 May 8.
Published in final edited form as: Cell Rep. 2014 May 1;7(3):722–734. doi: 10.1016/j.celrep.2014.04.025

Figure 7. CtBP binds to Esg and is required for hub maintenance.

Figure 7

(A) Western blot analysis from stable line extracts confirmed expression of EsgNLAP and NLAP with copper induction. Expected sizes of NLAP (33.3kDa) and EsgNLAP (85.3kDa). (B) Immunoprecipitation with anti-GFP antibodies and Western blot with anti-CtBP antibodies confirmed Esg and CtBP interaction. Expected size of CtBP (42.3 kDa). (C,D) Testes from 1-day old adult updGal4;UAS-CtBPRNAi;Gal80ts flies raised at 18°C (C) or shifted to 29°C for 10 days (D) immunostained for CtBP (green) and Fas3 (hub, red). Scale bars, 10μM. (E) Quantification of hub cell number in genotypes. Statistical significance shown with one-way ANOVA (Kruskal-Wallis test) and Dunn’s multiple comparison test (***p<0.001). (F) Frequency of hub cell-cyst cell conversion in control (updGal4;;G-TRACE/Gal80ts) or CtBPRNAi (updGal4; UAS-CtBPRNAi; G-TRACE/Gal80ts) flies. Testes with no detectable GFP+ cells at the 17 day-29°C time point were scored as negative. (G–H) Immunofluorescence images from control (G) or UAS-CtBPRNAi (H) flies raised and maintained at 18°C until 5 days after eclosion, shifted to 29°C for 7 days, and shifted back to 18°C. Testes immunostained for GFP (green), DsRed (red), and Fas3 (white). (I) Quantification of GFP+/dsRed− cells for the noted temperature regimes.