Genome-wide mapping reveals impaired initiation of DNA replication upon induced knockout of RAD51.
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(A) Graphs show the distribution of sites of DNA replication initiation across two complete chromosomes in the indicated cell lines, in each case grown in the absence (-RAP) or the presence (+RAP) of rapamycin; MFA-seq signal after cells were incubated with 5mM HU for 8 hours is also indicated. MFA-seq signal is represented by Z-scores across the chromosomes, calculated by comparing read depth coverage of DNA from exponentially growing cells relative to stationary cells; the bottom track for each chromosome displays coding sequences, with genes transcribed from right to left in red, and from left to right in blue. (B) Metaplots of MFA-seq signal found in every chromosome, centred on the previously mapped constitutive DNA replication origin (SSRORI) ±0.15 Mb, in -RAP and +RAP cells, and in the absence (-) or presence of HU (+). (C) Metaplots of MFA-seq signal across 0.15 Mb of sequence from all chromosomes ends in -RAP and +RAP cells, and with (+HU) or without (-HU) growth in the presence of HU. In B and C, p values were determined using Wilcoxon test by comparing -RAP with +RAP cells within each -HU and +HU pair.