Figure 5: RNA editing regulates heterochromatic gene silencing.
From: RNA editing regulates transposon-mediated heterochromatic gene silencing
(a) Representative images of control (dAdarLoxP) or dAdar null (dAdar0) male eyes expressing mini-white reporters of PEV located on chromosome 4, including two independent Hokmw alleles. Scale bar, 150 μm. (b) The mean pigmentation levels of the four reporters in control and dAdar0 eyes. n=4 independent replicates. (c) Western blots of silencing and activating chromatin marks in control and dAdar0 head samples. (d) Quantification of HP1 and histone methylation levels, normalized to Actin. n=7–12 western blots. (e) Confocal slices showing dimethyl H3K9 (H3K9me2) expression in control and dAdar0 adult neuronal nuclei. Scale bar, 5 μm. (f–h) Properties of H3K9me2 puncta in control and dAdar0 adult neuronal nuclei. n=30 nuclei/genotype. (i–j) Average linear signal intensities for DAPI, H3K9me2 and HP1 immunostaining across the neuronal nuclei in control (n=25 nuclei) and dAdar0 (n=20) adult brains (see Methods for details). Values were normalized to the peak DAPI signal. (k) Salivary gland nuclei labelled with antibodies against FIB and H3K9me2 (me2). Note the irregular nucleolar morphology in nuclei overexpressing dADAR (arrows, lower panel) relative to controls (upper panel). Scale bar, 20 μm. (l) Expression of a Lac-Z-based reporter of gene silencing (see Methods) in control (dAdarLoxP) and dAdarhyp oenocytes. n=30/genotype. (m–n) Lifespan curves for wild-type and dAdarhyp males in m and females in n (see Supplementary Table S1 for n-values and statistics). Error bars indicate s.e.m. *P<0.05, **P<0.005, ***P<0.0005, Mann–Whitney U-test.
