XB-IMG-151209
Xenbase Image ID: 151209
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Figure 8. Ki-67 is not required for rRNA biogenesis but controls gene transcription.(A) Northern-blot analysis of total RNA extracted from HeLa and U2OS cells constitutively expressing shRNA against Ki-67; and HeLa, U2OS, HCT-116 and HCT-116 TP53 (-/-) depleted of Ki-67 by siRNA for 72 hr in two biological replicates (#1 and #2) or with scrambled siRNA control (Scr). Pre-rRNA intermediates were analysed by probing with different primers located in the different spacers of the 47S sequence (5’ETS-green; ITS1-blue; ITS2-purple). (B) Quantification of 47S rRNA precursor in HeLa, HCT-116 and HCT-116 TP53 (-/-) depleted of Ki-67 by siRNA for 72 hr in three biological replicates (n=1–3). (C) U2OS cells (left) or HeLa cells (right) show transcriptome profile differences (fold change >1.5; corrected p-value <0.02) between asynchronous cells constitutively expressing control (CTRL) or Ki-67 shRNA. Heat-maps present the expression levels of differentially expressed genes between biological replicates (1,2,3) and technical replicates (3, 3*). Data is provided in Figure 8—source data 1 and 2.DOI:
http://dx.doi.org/10.7554/eLife.13722.03210.7554/eLife.13722.033Figure 8—source data 1. Ki-67-dependent transcriptome in U2OS cells.Table showing statistically significant (corrected p value < 0.02, Fold-change >1.5) changes of transcript abundance from Agilent Gene chip analysis of cDNA from control U2OS (pGIPZ-shRNA non silencing control) and U2OS stably silenced (pGIPZ-Ki-67 shRNA) for Ki-67.DOI:
http://dx.doi.org/10.7554/eLife.13722.033Figure 8—source data 2. Ki-67-dependent transcriptome in HeLa cells.Table showing statistically significant (corrected p value <0.02, Fold-change >1.5) changes of transcript abundance from Agilent Gene chip analysis of cDNA from control HeLa (pGIPZ-shRNA non silencing control) and HeLa stably silenced (pGIPZ-Ki-67 shRNA) for Ki-67.DOI:
http://dx.doi.org/10.7554/eLife.13722.034Figure 8—figure supplement 1. Ki-67 depletion does not hinder rRNA transcription.Newly synthesised RNA in asynchronous U2OS cells expressing control or Ki-67 shRNA visualised by 5-ethynyl uridine (EU) incorporation and immunofluorescence. Bar, 10 µm.DOI:
http://dx.doi.org/10.7554/eLife.13722.035 Image published in: Sobecki M et al. (2016) © 2016, Sobecki et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |