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. 2022 Jun 17;13:3493. doi: 10.1038/s41467-022-31211-9

Fig. 4. ZFP281 interacts with BRCA2.

Fig. 4

a Purification and mass spectrometry analyses of the ZFP281-associated proteins. Clonal cell lines expressing FLAG-tagged ZFP281 were generated in 293 Flp-in-TRex cells and the ZFP281 associated proteins were purified using the FLAG-affinity purification method and analyzed by SDS-PAGE, silver staining and mass spectrometry. At least two independent biological repeats with similar results. b Confirmation of the interaction of ZFP281 with EMSY, BRCA2, and QSER1 by endogenous immunoprecipitations. c Size exclusion chromatography of ES nuclear extracts demonstrated that the ZFP281, BRCA2, EMSY and QSER1 co-elute at ~ 2 MDa (fractions 9–12). d, e Different ZFP281 truncated proteins were expressed with an N-terminal FLAG tag in 293 T cells, and the interactions of BRCA2, EMSY, QSER1 with these ZFP281 truncated proteins were examined by FLAG immunoprecipitations, followed by western blot analyses. f, g Different BRCA2 truncated proteins were expressed with an N-terminal FLAG tag in 293 T cells, and the interactions of ZFP281, EMSY, QSER1 with these BRCA2 truncated proteins were examined by FLAG immunoprecipitations, followed by western blot analyses. At least three biological replicates were performed in b, c, e and g with similar results.