Fig. 6: DCAF13 mediates K63-linked ubiquitination at K1180 and K1184 sites on RPA194.

a GST pull-down assay using purified GST, GST-RPA194-N, GST-RPA194-M, GST-RPA194-C and DCAF13 protein in MDA-MB-231 cell lysates, and employing anti-GST beads as the bait, demonstrated direct binding region. b After transfection with FN-RPA194-WT, FN-RPA194-Nmut and FN-RPA194-Cmut plasmid respectively, the Hela cells were directly subjected to IF experiments. The subcellular localization and morphology of the different forms of RPA194 were then observed under confocal microscopy. c After transfecting the RPA194 single point mutant plasmid and HA-K63 in Hela cells, the ubiquitination levels of different mutant form of RPA194 were analyzed by western blot. d The spatial structure of the Pol I complex mapped by ChimeraX, where K1101, K1180, K1184 and K1574 are located on the periphery of the complex. e Western blot analysis of the effect about over-expression of DCAF13 on K63-linked ubiquitination of RPA194 wild type and mutant. f IF assay of EGFP-FBL knock-in Hela cells after transfected with Ruby3-RPA194-WT or Ruby3-RPA194-K1180/1184 R. Scale bars, 5 μm (b, f). Source data are provided as a Source Data file.