Fig. 7: DCAF13-mediated K63-linked ubiquitination of RPA194 promotes the transcriptional activity of Pol I. a.

Western blot analysis of RPA194 levels in MDA-MB-231 cells transfected with either scramble control or siDCAF13 and subsequently treated with 10 μg/mL cycloheximide (CHX) for specified durations. b Western blot analyses detecting POLR1B, POLR1C, POLR1D, and POLR1E levels in MDA-MB-231 cells transfected with either scramble control or siDCAF13, following immunoprecipitation with an anti-POLR1A antibody. c Chromatin immunoprecipitation sequencing (ChIP-Seq) assay identifying RPA194 enrichment on rDNA in MDA-MB-231 cells transfected with either scramble control or siDCAF13. Read normalization was accomplished using Counts Per Million (CPM) via deepTools; the structure of 18S, 5.8S, and 28S rRNA genes is depicted below the profile. d ChIP-qPCR analysis detecting the binding of TAF1C to rDNA in MDA-MB-231 cells transfected with scramble control or siDCAF13 (n = 3 biological replicates). e ChIP-qPCR analysis detecting the binding of Pol I to rDNA in Hela cells transfected with EV, FN-RPA194-WT or FN-RPA194-K1180/1184 R (n = 3 biological replicates). f Western blot analysis of the levels of K63-linked ubiquitination of RPA194 in HeLa cells transfected with either scramble control or siCK2. g qRT-PCR assays measuring RNA levels of CK2 and pre-rRNA in HeLa cells transfected with siCK2 (n = 3 biological replicates). h Western blot analysis of the levels of K63-linked ubiquitination of RPA194 in HeLa cells transfected with an oe-DCAF13 plasmid and/or siCK2. d, e, g Data shown as mean ± SEM. Statistical significance was calculated by two-tailed unpaired Student’s t-test. Source data are provided as a Source Data file.