IMAGE

Figure 2

ID
ZDB-IMAGE-220525-23
Source
Figures for Idilli et al., 2020
Image
Figure Caption

Figure 2

Rad21 depletion prevents ALT, normalize telomere length and reduces TERRA levels in brain tumors. (A) Telomere length analysis through telomeric qPCR in RAS, RAS Rad21 +/− brain tumors compared to control brains (dashed lines). The relative amount was normalized to the signal of a single copy gene (rps11). Each dot represents one sample, n = 3 in all groups. (B) Representative fluorescent microscope images of Q-FISH analysis of RAS and rad21;RAS nuclei with ultrabright foci (white arrows). Scale bar: 5 µm. (C) Quantification of the numbers of telomere foci per nucleus and (D) quantification of relative telomere length intensity measured by Q-FISH in RAS and rad21;RAS brain tumors. * p < 0.05, ** p < 0.01. RAS n = 48 nucleus; rad21;Ras n = 46 nucleus. (E) Analysis of coefficient of variation of telomere mean intensity measured in (C) between RAS and rad21;RAS. (F) Distribution of telomere length evaluated by Q-FISH (C) in RAS and rad21;RAS tumors. The signals >2700 AU could represent ultra-long telomeres or telomeric clusters, an ALT feature. (G) Quantification analysis of C-circle assay dot blots (each dot represents one sample). Determination of C-circles amount was calculated after subtracting global background and specific –θ29 signal. AU: arbitrary unit. (H) Representative C-circle assay followed by telomeric dot blot in control, RAS and rad21;RAS brain tumors compared with telomerase+ HeLa cells and ALT+ U2OS cells. Reactions without phy29 polymerase (–θ29) were included as a control. (I) Quantification of TERRA expression measured by dot blot using total RNA (500 ng) of control, RAS, and rad21;RAS brain tumors. As positive and negative control, U2OS and Hela cells were added. (J) Representative RNA dot blot hybridization using total RNA (500 ng) of control, RAS, and rad21;RAS brain tumors. As positive and negative control, U2OS and Hela cells were added. For (G,I), a minimum of three independent experiments were performed and analyzed with 1 way anova Kruskal–Wallis—non-parametric test.

Figure Data
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Genes (Basel)