Fig. 1: Loss of RIOK2 results in telomere shortening.
From: RIOK2 transcriptionally regulates TRiC and dyskerin complexes to prevent telomere shortening
a Immunoblot showing knockdown (KD) and knockout (KO) of RIOK2 in TF-1 cells using 2 guide RNAs each: KD#1, KD#2 and KO#1, KO#2. 2 different replicates for KD#2 (KD#2.1, 2.2) and KO#2 (KO#2.1, 2.2) are shown. b Proliferation in TF-1 cells after KD and KO of RIOK2 using 2 guide RNAs each: KD#1, KD#2 and KO#1, KO#2; n = 2 independent experiments run simultaneously and plotted together. c Cell cycle analysis of control (Ctrl) vs RIOK2 KD and KO TF-1 cells after days 4, 6, and 8 of gene-editing; n = 3 experimental replicates. d Gene set enrichment analysis (GSEA) plot of telomere maintenance-associated genes from RNA-sequencing in RIOK2-depleted vs control HSPCs. e GSEA plot of telomere maintenance-associated genes from ATAC-sequencing in RIOK2-depleted vs control HSPCs. Chromatin accessibility at the promoters of genes were analyzed and GSEA analyses performed. f, g, h qPCR-based analysis of telomere lengths in HSPCs, TF-1 and K562 cells respectively, upon RIOK2 deficiency. Equal amounts of genomic DNA (gDNA) were loaded across samples to assess telomere lengths/content by measuring T/S ratio (telomere/single copy gene expression), as previously described34,35; n = 3 primary human donors in figure f and n = 3 experimental replicates in figures g and h. i, j Fluorescence in-situ hybridization (FISH) of telomeric DNA repeats (TTAGGG) upon RIOK2 deficiency in TF-1 and K562 cells, respectively. Representative images from 3 experimental replicates. Scale bar 10 µm. k, l qPCR-based analysis of telomere lengths in control vs RIOK2-depleted HeLa and HEK293 cells, respectively. n = 3 experimental replicates. m Schematic illustration of assessment performed on MDS patient-derived bone marrow (BM) cells. n Correlation of the mRNA expression of RIOK2 with telomere lengths in MDS patient-derived BM cells; n = 40. AU: arbitrary unit. One-way analysis of variance (ANOVA) with Dunnett’s multiple comparisons test performed in c, f, g, h, k and l. Data represented as mean ± SEM. All comparisons are done w.r.t. control (Ctrl). Two-tailed nonparametric Spearman correlation performed in n and Spearman correlation coefficient (r) and P values are shown. ns: non-significant. See also Supplementary Fig. 1. Source data are provided as a Source Data file.
