Fig. 4: TER94 influences Mu2 level in regulating nuclear size.
a Quantification of the R1-R6 cross-section nuclear area from Rh1 > TER94K2A flies co-expressing UAS-RNAi constructs targeting indicated genes. The purple lines indicate the difference of the mean equals 1 (mean diff. = 1) as compared to the control (Rh1 > TER94K2A, LacZ). The DDR genes with mean diff. ≥1 are marked in yellow. The number of independent nuclei measured are listed in Statistics and reproducibility. b Confocal and 3D rendering images (individual nuclei are color-coded) of 7-day-old (confocal) or 5-day-old (3D rendering) retinas from flies expressing indicated transgenes stained with phalloidin (red) and anti-Lamin (magenta). c–e Quantification of the cross-section area of R1-R6 nuclei (c), eGFP intensity (d), and nuclear volume (e). f Confocal images of 7-day-old retinas expressing indicated transgenes stained with anti-Lamin (red) and anti-γH2AV (magenta). g Quantification of the anti-γH2AV intensity. h Pearson’s correlation of nuclear size and eGFP-Mu2 intensity (r: coefficient). i Western analysis of eGFP-Mu2 level (anti-GFP) in indicated TER94 backgrounds. Anti-Histone H3 and anti-β-Actin serve as loading controls. j RT-PCR measurement of mu2 and TER94 expression from indicated genotypes. The control (Ctrl) groups omitted templates. rp49 serves as control. k Western analysis of Mu2 level in tissues expressing LacZ or TER94K2A. Anti-Histone H3 and anti-β-Actin serve as loading controls. l Larval eye discs from flies expressing indicated transgenes under the control of GMR-GAL4 are stained with anti-GFP (magenta) and anti-V5 (red) (left panel). Larvae bearing the same genotypes are subjected to in situ PLA using anti-GFP and anti-V5 antibodies (right panel). The duolink signal reveals proteins in the same complex. The lines in dot plots from a represent mean ± SE. The number of independent experiments performed: 3 (b, i), 2 (j, k, l). Quantification details of c, d, e, and g are listed in Statistics and reproducibility. Scale bars: 10 µm (b, f), 20 µm (l).
