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. Author manuscript; available in PMC: 2021 Jan 15.
Published in final edited form as: Nature. 2020 Jul 15;585(7824):298–302. doi: 10.1038/s41586-020-2497-0

Extended data Fig. 5 |. Artificial and natural modulation of sincRNA levels.

Extended data Fig. 5 |

(a) In HEK293 cells treated with the Pol II inhibitor flavopirodol (FP), introduction of antisense oligonucleotides (ASO) targeting sincRNAs does lower IGS ncRNA levels relative to ASO control-treated cells (CTL). ASO-dependent percent decreases in sincRNA levels are indicated for each IGS site and the average decrease in total sincRNA levels is 49%. Data are shown as the mean±s.d.; two-tailed t-test, n=3 biologically independent experiments. (b-d) In the absence of heat shock, artificial overexpression of sincRNA22 (psincRNA) in the nucleolar stress hyper-responsive MCF7 cells failed to repress rRNA biogenesis (b) or rRNA levels (c) despite the enrichment of the sincRNA22 in the nucleolar fraction (d). Plasmid (pCTL), iPol I (LAD), vehicle (DMSO), and GAPDH cell fractionation controls were included. Data are shown as the mean±s.d.; n=2 biologically independent experiments (b, d); two-tailed t-test, n=3 biologically independent experiments (c); (e) Quantification of the number of distinct NPM foci per cell in different cell types. Data are shown as the mean±s.d.; one-way ANOVA with Tukey’s multiple comparisons test, n=5 biologically independent experiments.