Fig. 1
From: Nuclear lamina integrity is required for proper spatial organization of chromatin in Drosophila
Chromatin is released from the NE and becomes denser upon Lam-KD in S2 cells. a, b Western-blot analysis of lamin Dm0 and lamin C protein levels in Drosophila cell lines (a), or in Lam-KD and control S2 cells (b). Band intensity quantitation is presented below. c A representative example of nuclei immunostained with antibodies against histone H4 and LBR. Fluorescence intensity along the yellow-framed zone was measured using ImageJ software. d Averaged fluorescence intensity profiles along the nuclear diameter in Lam-KD (n = 120) and control (n = 180) S2 cells immunostained with antibodies against histone H4 and LBR. P value for the framed region of histone H4 profiles was estimated in a Wilcoxon test. e Cumulative frequency of radial positions of the FISH probe located in the LAD from cytological region 36C in Lam-KD (n = 175) or control (n = 175) S2 cells (right panel). P value was estimated in a Kolmogorov-Smirnov one-sided test. Representative examples of FISH signals in nuclei stained with anti-LBR antibody are shown on the left panels. Scale bar 1 µm. f Cumulative frequency of radial positions of FISH probes to cytological regions 22A (left) or 60D (right) in Lam-KD (n = 100 for 22A and n = 99 for 60D) or control (n = 115 for 22A and n = 72 for 60D) S2 cells. P value was estimated in a Kolmogorov-Smirnov one-sided test. Data for analysis were taken from ref. 11. g Total chromatin volume measured by DAPI fluorescence in Lam-KD (n = 125) or control (n = 83) S2 cells. P value was estimated in a Wilcoxon test. Thick black lines and white dots represent median and average values, upper and lower ends of boxplot show the upper and lower quartiles, the whiskers indicate the maximum and minimum values. h Averaged fluorescence intensity profiles along the nuclear diameter in LBR-depleted (LBR-KD, n = 120) or control (n = 120) S2 cells immunostained with antibodies against histone H4 and lamin Dm0. NS – non-significant difference (P > 0.05, Wilcoxon test). i Cumulative frequency of radial positions of the FISH probe to cytological region 36C in LBR-KD (n = 150) or control (n = 150) S2 cells. NS – non-significant difference (P > 0.05, Kolmogorov-Smirnov one-sided test)
