Fig. 3 | Chromosoma

Fig. 3

From: Ectopic expression of pericentric HSATII RNA results in nuclear RNA accumulation, MeCP2 recruitment, and cell division defects

Fig. 3

HSATII RNA accumulates adjacent to site of integration and recruits MeCP2. (a) RNA hyb for HSATII RNA (red) followed by sequential DNA hyb using a probe to detect the pTargeT backbone (green) demonstrates HSATII RNA accumulates primarily adjacent to sites of pTargeT genome integration in HeLa cells stably expressing HSATII 3 weeks after transfection. All boxes with white border indicate regions of interest with zoomed image at bottom right. Individual channels images shown in black and white. (b) Scatter plot of Pearson correlation coefficients for individual nuclei (n=10) hybridized for sequential HSATII RNA/pTargeT DNA and HSATII RNA with MeCP2 co-IF (HSATII RNA/MeCP2). Black lines indicate mean; grey error bars indicate 95% confidence interval. (c) Co-HSATII RNA hyb and IF for MeCP2 indicate a near perfect colocalization of HSATII RNA and MeCP2 signals within HeLa nuclei 3 weeks following transfection, as demonstrated by Pearson correlation coefficients plotted in (b). (d) Dual channel linescan of inset shown in (c) demonstrating colocalization of MeCP2 (red) and HSATII RNA (green). (e) RNA hyb for HSATII RNA (red) followed by sequential DNA hyb using a probe to detect the pTargeT backbone (green) demonstrates HSATII RNA accumulates adjacent to, and not overlapping, sites of genome integration in Tig-1 cells stably expressing HSATII 3 weeks after transfection. (f) Dual channel linescan of inset in (e) demonstrating the HSATII RNA (red) and pTargeT DNA (green) signals are adjacent, and not overlapping. (g) Tig-1 fibroblasts stably expressing HSATII RNA for 3 weeks demonstrate overlapping signals for HSATII RNA and MeCP2. Box with white border indicates region of interest with zoomed image at bottom right. (h) Dual channel linescan of inset shown in (g) demonstrating colocalization of MeCP2 (red) and HSATII RNA (green). Scale bar, 5μm

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