Fig. 5: p53 acts downstream of Mu2 in TER94 dysfunction-induced nuclear expansion. | Nature Communications

Fig. 5: p53 acts downstream of Mu2 in TER94 dysfunction-induced nuclear expansion.

From: VCP maintains nuclear size by regulating the DNA damage-associated MDC1–p53–autophagy axis in Drosophila

Fig. 5

a Left: Confocal micrographs of 5-day-old control (Rh1 > LacZ) or Rh1 > Mu2 adult retinas co-expressing LacZ or p53-RNAi stained with phalloidin (red) and anti-Lamin (magenta). Right: Quantification of the cross-section area of R1-R6 nuclei. b Left: Confocal images of 6-day-old Rh1 > TER94K2A adult retinas in wild type (+/+) or p53 null (−/−) background stained with phalloidin (red) and anti-Lamin (green) antibody. Right: Quantification of the cross-section area of R1-R6 nuclei. c Left: Confocal and 3D rendering images (individual nuclei are color-coded) of 5-day-old Rh1 > eGFP-Mu2 adult retinas in wild type (+/+), p53 null (−/−), 6XMyc-p53A overexpression in p53 null (−/−), and 6XMyc-p53B overexpression in p53 null (−/−) backgrounds stained with phalloidin (red) and anti-Lamin (magenta). Right: Quantification of the cross-section area and the volume of R1-R6 nuclei. d Western analysis of p53 level in tissues expressing LacZ (control) or TER94K2A under the control of Rh1-GAL4. The arrows indicate molecular weight of each p53 isoforms. Asterisk: nonspecific. Two independent experiments showed similar results. e Western analysis of the Myc-p53A level in tissues expressing indicated genes. Anti-Histone H3 and anti-β-Actin serve as loading controls. f Larval eye discs expressing indicated genes are stained with anti-GFP (magenta) and anti-Myc (red) antibodies (left panel). Larvae bearing the same genotypes are subjected to in situ PLA using anti-GFP and anti-Myc antibodies (right panel). The duolink signal reveals proteins in the same complex. g Top: Confocal micrographs of 5-day-old Rh1 > LacZ, Rh1 > 6XMyc-p53A > LacZ, and Rh1 > 6XMyc-p53A > TER94K2A retinas stained with phalloidin (red), anti-Lamin (magenta), and anti-Myc (green). Bottom: Quantification of the cross-section area of R1-R6 nuclei. The number of independent experiments performed: 3 (b), 2 (f). Quantification details of a, b, c, and g are listed in Statistics and reproducibility. Scale bars: 10 µm (a–c, g), 20 µm (f).

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