Fig. 4: DCAF13 promotes global translation by interacting with Pol I.
a Overexpression of DCAF13 promotes the synthesis rate of nascent proteins in MDA-MB-231 and MDA-MB-468 cells. b Silver staining assay identifying proteins interacting with DCAF13 following Immunoprecipitation with anti-DCAF13 antibody. c Complex enrichment analysis of DCAF13-interacting proteins, with log10 p-value calculated using Benjamini linear step-up correction method. d IF assays detecting the localization of DCAF13 and RPA194 in MDA-MB-231 cells treated with DMSO, ActD, or transfected with siRNA targeting to DCAF13. e IF assays of full-length DCAF13 or truncated DCAF13 using anti-HA antibody to visualize its localization. f 5-Ethynyl uridine (EU) assays of MDA-MB-231 cells transfected with either a scramble or siDCAF13. g, h qRT-PCR assay detecting the RNA levels of DCAF13 and nascent pre-rRNA after knocking down (siD13) (g) or overexpressing (oeD13) (h) DCAF13 in MDA-MB-231 cells (n = 3 biological replicates). i IF assays in MDA-MB-231 cells treated with DMSO, ActD, or transfected with siDCAF13, siDDB1, and siCUL4A, with simultaneously stained for UBF and NPM. j RNA-fluorescence in situ hybridization (RNA FISH) assay of MDA-MB-231 cells treated with DMSO or ActD using a Cy3-U3 DNA probe, co-staining with NPM and UBF antibodies, to label U3 and nucleolar markers concurrently. k-l Co-IP assay of MDA-MB-231 cells using either anti-IgG, anti-DCAF13 (k), or anti-RPA194 antibody (l), and analyzed by Western blotting with indicated antibodies. Scale bars, 5μm (d–f, i, j). g, h 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.
