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. 2015 Jan 13;43(3):1659–1670. doi: 10.1093/nar/gku1406

Figure 5.

Figure 5.

53BP1 inhibits MMEJ and end-resection/deletion in the BRCA1-deficient cells. (A) Protein levels of 53BP1 in H1299 cells with or without BRCA1 depletion were detected by western blot using an anti-BRCA1/53BP1 antibody or β-actin as a protein loading control. (B and C) MMEJ measured by the extrachromosomal or chromatinized reporter pCMV/myc/cyto/GFP* in G1-enriched cells, according to the protocol described in Figure 3A, P-values were calculated by Student's t-test. Error bars represent the SD of three independent experiments. (D) Quantitative analysis of deletion products by sequence analysis of individual re-circularized pGL3-MCS in cells depleted of the indicated genes. (E) HA-I-SceI expression and BRCA1/53BP1 expression are monitored by western blot. (F) Quantitation of changes in NHEJ events. A cell clone with a single copy of pPHW1 (H1299-pPHW1) was enriched in G1 phase using starvation as described in Figure 3A. To avoid selection bias, colonies did not undergo XHATM selection for gpt expression. P < 0.05, P-values were calculated by Chi-square test.