Fig. 6: RTE expression and activity are upregulated in Cap-d3 insufficient larval neuroblasts.

a qRT-PCR analyses of RTE (mdg1, mdg4, and X-element) transcripts were performed with cDNA generated from third instar larval brains dissected from w¹¹¹⁸ controls or cap-d3 transheterozygous mutant larvae (150 brains per genotype; mixed sexes). Transcripts were normalized to the housekeeping gene, rp49. Results show the averages of three independent biological replicates. P values were determined by performing two-tailed, unpaired t-tests. For mdg1, **p = 0.0054, for mdg4, **p = 0.0015, and for x-element, **p = 0.0067. b Third instar larval brains were dissected from male larvae expressing UAS-gypsyCLEVR⁵¹ and UAS-GFP dsRNA or UAS-cap-d3 dsRNA under the control of eyGAL4, GMRGAL4, immunostained with antibodies to mCherry, a reporter of gypsy retrotransposition (pseudocolored green), and stem cell marker, Dpn (red), and imaged using confocal microscopy. Maximum projections of z-stacks are shown. Nuclei are stained with DAPI (blue). All images were taken with 40x magnification. Scale bar = 50 µm. c The percentage of cells staining positive for both mCherry and Dpn were quantified from experiments described in (b). Male larval brains expressing eyGAL4 and GMRGAL4-driven UAS-gypsyCLEVR were also analyzed as “no RNAi” controls. Results shown include larvae harvested from two independent experiments; each data point represents a single brain. d Third instar larval brains were dissected from male larvae expressing UAS-gypsyCLEVR and control UAS-GFP dsRNA (green circles) or UAS-cap-d3 dsRNA (orange squares), under the control of eyGAL4, GMRGAL4, immunostained with antibodies to detect mCherry and Dpn, and imaged using confocal microscopy. The percentage of cells that stained positive for both Dpn and Dcp1 were quantified. Flies were developed on food containing DMSO as a control (closed circles and squares), or on food containing 5 µM AZT (open circles and squares). Results shown include larvae harvested from two independent experiments; each data point represents a single brain. For experiments in (c, d), P values were determined by performing two-tailed Mann–Whitney analyses. For (c), **p = 0.0029, ****p ≤ 0.0001, NS not significant. For (d), **p = 0.0037, **#p = 0.0039, NS not significant. Error bars indicate standard deviations from the mean.