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. 2022 Aug 30;11:e75879. doi: 10.7554/eLife.75879

Figure 11. Cell cycle progression is required for the precise timing of Slp expression and this is mediated in part through Notch signaling.

(A-A’’’) The expression of PCNA, Dpn (red), and E(spl)mγGFP (cyan) in vsx-Gal4 driving UAS-DCR2 and UAS-PCNA-RNAi (VDRC51253) optic lobes (n=5). The affected region is indicated by loss of PCNA staining. (B-B’’’) The expression of Dpn and SoxN in vsx-Gal4 driving UAS-DCR2 and UAS-PCNA-RNAi optic lobes (n=5). (C-D’’’) The expression of Dpn, Ey, and Slp2 in vsx-Gal4 driving UAS-DCR2 and UAS-PCNA-RNAi optic lobes (n=9) showing the surface neuroblast layer view (C-C’’’’) and the deep progeny layer view (D-D’’’) of the same brain. (C-C’’’’) In affected region marked by loss of PCNA staining, Ey expression is reduced and Slp2 expression is lost in Dpn +neuroblasts. Some glia express Slp2 but not Dpn are indicated by white arrows. (D-D’’’) Dpn expressing neuroblasts are observed in deep layers in the affected region, but they don’t express Ey or Slp2. (E-E’’’) Larvae of genotype ywhsFLP; ay-Gal4 UAS-GFP /UAS-PCNA-RNAi; UAS-DCR2/+ were heat shocked at 37°C for 8 min 70 hours before dissection at 3rd instar larval stage. Ey expression (blue) is minimally affected in clones marked by GFP (green), while Slp2 (red) expression is delayed (n=17 clones). (F-F’’) Magnified view of the boxed region in E’’’. (G-G’’’) Larvae of genotype ywhsFLP; ay-Gal4 UA-SGFP /UAS-PCNA-RNAi; UAS-DCR2/dpn >FRT-STOP-FRT-NICD were heat shocked at 37 °C for 8 min 70 hr before dissection at 3rd instar larval stage. Ey expression (blue) is minimally affected in clones marked by GFP (green), while Slp2 (red) expression initiation is rescued (n=10 clones). (H-H’’) Magnified view of the boxed region in G’’’. (I-J’’’) The expression of Ey, Slp2, and Dpn in ywhsFLP; dpn >FRT-STOP-FRT-NICD (I-I’’’) and yw control (J-J’’’) optic lobes (n=5 each) (heat shocked 37°C for 8 min 70 hr before dissection). Scale bars 20 µm.

Figure 11.

Figure 11—figure supplement 1. Cell cycle progression is required for the temporal cascade progression.

Figure 11—figure supplement 1.

(A-A’’’) Larvae of genotype ywhsFLP; ay-Gal4 UAS-GFP /+; UAS-DCR2/UAS-stgRNAi (VDRC 17760) were heat shocked at 37°C for 8 min 70 hr before dissection at 3rd instar larval stage. Exd expression (co-factor of Hth, and always expressed in the same pattern) is normal in clones marked by GFP (gray), Ey expression is slightly delayed, while Slp2 expression is severely delayed (11 clones). (B-B’’’) Magnified view of the boxed region in A’’’’. (C-C’’’’) Larvae of genotype ywhsFLP; ay-Gal4 UAS-GFP /+; UAS-DCR2/UAS-Dap (BDSC 83338) were heat shocked at 37°C for 8 min 70 hr before dissection at 3rd instar larval stage. Exd expression is normal in clones marked by GFP (gray), Ey expression is slightly delayed, while Slp2 expression is severely delayed (8 clones). (D-D’’) Magnified view of the boxed region in C’’’’.