Fig. 3: Blocking TER94 function hampers DNA damage repair. | Nature Communications

Fig. 3: Blocking TER94 function hampers DNA damage repair.

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

Fig. 3

a Confocal images of 5-day-old Rh1 > LacZ, Rh1 > Derlin-1, and Rh1 > Derlin-1ΔSHP retinas stained with phalloidin (red), anti-Lamin (green), and anti-TER94 (magenta). b Left: Analysis of TER94 subcellular distribution. The analyzed range is indicated by the white lines shown in panel a. Image J measures the intensities of anti-Lamin and anti-TER94 signals along the lines. The peaks of the anti-Lamin signal define the borders of the nuclei. Right: Quantification of the cross-section area of R1-R6 nuclei (top) and the nuclear TER94 intensity (cumulated pixel of TER94 along the line divided by the line distance defined by anti-Lamin-marked two peaks, bottom). c Confocal images of wild-type larval eye discs with or without 50 µM H2O2 treatment stained with anti-Lamin (red) and anti-γH2AV (green). d Time-course analysis of the change in nuclear size and the level of γH2AV from Rh1 > LacZ and Rh1 > TER94K2A adult eyes stained with phalloidin (red), anti-Lamin (magenta), and anti-γH2AV (green). e Quantification of the anti-γH2AV signal intensity over time from Rh1 > LacZ and Rh1 > TER94K2A. f An adult TER94K15502 clone, marked by the absence of myr.GFP (arrowheads), stained with anti-Lamin (magenta) and anti-γH2AV (red). R cell boundaries are outlined in dash lines. g Confocal images of Rh1 > LacZ and Rh1 > TER94K2A adult retinas stained with phalloidin (red), anti-Lamin (magenta), and anti-PCNA (green). h Left: Confocal images of 5-day-old Rh1 > TER94K2A retinas from flies raised in normal 12 h light/12 h dark (L/D) or complete dark (D/D) conditions after birth stained with phalloidin (red), anti-Lamin (magenta), and anti-γH2AV (green). Right: Quantification of the γH2AV intensity (top) and the cross-section area of R1-R6 nuclei (bottom). i Left: Confocal images of 5-day-old retinas from Rh1 > TER94K2A co-expressing either LacZ or hSOD1 stained with phalloidin (red), anti-Lamin (magenta), and anti-γH2AV (green). Right: Quantification of the γH2AV intensity (top) and the cross-section area of R1-R6 nuclei (bottom). The number of independent experiments performed: 2 (a), 4 (c), 6 (f), 4 (g). Quantification details of b, e, h, and i are listed in Statistics and reproducibility. Scale bars: 5 µm (a, d, f), 20 µm (c), 10 µm (g, h, i).

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