Figure 4. PALB2 directly interacts with RNF168 via its WD40 domain.
(A) Pulldowns of the indicated GFP fusion proteins in U2OS cells. Blots were probed for PALB2 and GFP. (B) In vitro GST pulldown of GST-PALB2 or GST alone in the presence of His-RNF168. Blots were probed for GST and His. (C) Schematic representation of full-length PALB2 (WT), five non-overlapping fragments (T1–T5) spanning PALB2, and two deletion mutants lacking the ChAM (T6) or WD40 domain (T7). (D) As in B, but using the indicated GST-tagged PALB2 fragments. (E) mCherry-RNF168 pulldowns from U2OS cells expressing the indicated YFP- tagged fusion proteins. Blots were probed for mCherry and YFP. (F) Pulldowns of the indicated YFP fusion proteins in U2OS cells either with or without expression of mCherry-RNF168. Blots were probed for GFP, mCherry and RAD51. (G) Pulldowns of the indicated GFP fusion proteins in U2OS cells. Blots were probed for PALB2, BRCA2 and GFP. (H) In vitro GST pulldown of GST-PALB2 or GST alone in the presence of His-RNF168 and/or His-piBRCA2. Blots were probed for GST and His.
Figure 4—figure supplement 1. RNF168 and BRCA2 interact with PALB2 in a non-mutually exclusive manner. .
