Figure 4
From: Noncoding human Y RNAs are overexpressed in tumours and required for cell proliferation
RNA interference against hY RNAs. (A) Quantification of RNA levels after RNAi. Three small interfering RNAs (designated as siRNAs a, b and c) directed against hY1, hY3, hY4 and hY5 RNA were transfected into proliferating HeLa cells. RNA was isolated at 48 h after transfection and the amounts of each targeted hY RNA relative to a calibrator RNA were determined by qRT–PCR. 5.8S rRNA was used as calibrator for hY1, and HPRT mRNA for the other hY RNAs. The expression of each target hY RNA after the experimental RNAi is shown as the percentage of the relative expression levels observed after a control RNAi against nontarget firefly luciferase mRNA. Mean values are shown for n independent experiments as indicated. (B) Quantification of replicating S phase cells after RNAi. At 47 h after transfection of asynchronously proliferating HeLa cells with the indicated siRNAs, replicating cells in the population were labelled for 1 h with BrdU. At 48 h, percentages of S phase cells incorporating BrdU into their chromosomal DNA were determined by immunofluorescence microscopy. Mean values and s.d. are shown for n independent experiments.
