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. 2021 Dec 13;17(12):e1009609. doi: 10.1371/journal.pgen.1009609

Fig 3. Lineage-wide PI3K/Tor hyperactivation does not induce CySC loss.

Fig 3

Control (A,B,G), Dp110 over-expression (C,D,H), or Tsc1 knockdown (E,F,I) in the somatic lineage using tj-Gal4 (A,C,E,G-H) or C587-Gal4 (B,D,F). A,C,E. Zfh1 (magenta) labels CySCs and early daughters, Eya and Fas3 (white) label differentiated cyst cells and the hub, respectively and germ cells are labelled with Vasa (cyan). B,D,E. Esg-GFP expression is shown in yellow, Tj (magenta) labels CySCs and differentiating cyst cells and Eya and Fas3 are shown in cyan. Zfh1- or Esg-expressing CySCs are observed upon PI3K or Tor hyperactivation in all conditions. G-I. p4E-BP staining (yellow) is increased upon Dp110 over-expression (H) or Tsc1 knockdown (I). Tj labels the somatic lineage (magenta). J. Graph showing the number of Zfh1-positive, Eya-negative CySCs upon Tor pathway hyperactivation. No significant changes were observed (Kruskal-Wallis, P<0.36, N = 19 for Tj>+, N = 18 for Tj>Dp110 and N = 11 for Tj>Tsc1 RNAi). K. Quantification of p4E-BP fluorescence intensity in CySCs surrounding the hub. Over-expression of Dp110 or knockdown of Tsc1 significantly increases p4E-BP intensity (Kruskal-Wallis and Dunn’s multiple comparisons, P<0.0003 for Tj>Dp110 vs Tj>+, P<0.0001 for Tj>Tsc1 RNAi vs Tj>+, N = 43 for Tj>+, N = 33 for Tj>Dp110 and N = 32 for Tj>Tsc1 RNAi). Lines in J and K indicate mean and 95% confidence interval. The hub is outlined with a blue dotted line. Scale bar in panel A panels represents 20 μm for A-F and scale bar in panel G panels represents 20 μm for G-I.