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. 2021 Oct 13;12:5966. doi: 10.1038/s41467-021-26227-6

Fig. 4. Identification of MCM10 as a BRCA2-ineracting protein.

Fig. 4

a Numbers of tryptic peptides obtained from two independent PALB2 tandem affinity purification (TAP)-LC-MS/MS experiments. b Co-IP of endogenous PALB2 and BRCA2 with ectopic MCM10. FLAG-HA-tagged MCM10 was transiently overexpressed in 293T cells and IPed with anti-FLAG. c Co-IP of endogenous MCM10 and endogenous PALB2 and BRCA2 in unperturbed and HU-treated U2OS cells. d Co-IP of GFP-tagged BRCA2 variants and FLAG-HA-tagged MCM10 overexpressed in 293T cells. e Association of endogenous BRCA2 with ectopic FLAG-HA-tagged MCM10. The procedure was performed as in (b) except that the final IP material was treated with DNase I on the beads and then washed before being subjected to western blotting. f Co-IP of BRCA2 and MCM10 in S-phase-synchronized U2OS cells at 3 h after treatment with DMSO, 10 Gy of IR, 2 Gy of IR, BLEO (1 µM), CPT (1 µM), or cisplatin (CDDP, 1 µM). Cells were synchronized in the S phase by a single thymidine block for 24 h and released for 2 h prior to the treatments. For (b–f), similar results were obtained in n = 3 independent experiments. Source data are provided as a Source Data file.