Fig. 5: Interactions of DONSON and FANCM with replisomes and chromatin in nontreated cells. | Nature Communications

Fig. 5: Interactions of DONSON and FANCM with replisomes and chromatin in nontreated cells.

From: DONSON and FANCM associate with different replisomes distinguished by replication timing and chromatin domain

Fig. 5

a DONSON associates with some, but not all, replisomes in untreated cells. Chromatin was prepared from untreated GFP-DONSON-HeLa cells and sequential IP performed, first against GFP-DONSON, and then against the GINS protein PSF1 from the residual supernatant. Representative blot (n = 3). b PLA of GFP-DONSON and PSF1 demonstrates DONSON-associated replisomes are more frequent in early S phase than in late S phase in NT cells. Scored nuclei: PLA between GFP-D: PSF1 early S phase = 82, late S phase = 81, from three biological replicates. Data are mean ± s.e.m. c PLA between FANCM and MCM2 demonstrates low level of FANCM-associated replisomes in late S phase in nontreated cells. Scored nuclei: PLA between FANCM: MCM2 of early S phase = 73, late S phase = 75, from three biological replicates. Data are mean ± s.e.m. d IP of FANCM demonstrates low-level interaction with replisome protein MCM2. e PLA between GFP-DONSON and H3K4me3 or H3K9me3. Scored nuclei of GFP-DONSON and H3K4me3 in early S phase = 79, late S phase = 78. Scored nuclei of GFP-DONSON and H3K9me3 in early S phase = 77, late S phase = 82, from three biological replicates. Data are mean ± s.e.m. f PLA between FANCM and H3K4me3 or H3K9me3. Scored nuclei of FANCM and H3K4me3 in early S phase = 64, late S phase = 65. Scored nuclei of FANCM and H3K9me3 in early S phase = 68, late S phase =  65, from three biological replicates. Data are mean ± s.e.m. For PLA experiments in b, c, e, f, a two-sided Mann–Whitney rank-sum test was used to determine if differences were statistically significant. NS: not significant: P > 0.05. Source data are provided as a Source Data file.

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