Fig. 5

Deep sequencing analysis of MED12 exon 2 mutations in primary myometrial cells with oxidative exposure in vitro. a Diagram illustrated the PQ treatment in primary myometrial cells, oxidation of DNA change, abasic (AP) DNA damage, and potential misrepair detected by deep sequencing for MED12 exon 2 in a short- (acute) and long-term (chronic) PQ treatment. b-e. (b) Counts of MED12 point mutations in four LM with known MED12 mutations (positive controls) originally identified by Sanger sequencing (positive controls of c.122, c.128, c.130, and c.130 mutations with detected mutation rates of 58.1, 25.0, 33.3 and 22.7% in the number and distribution of MED12 mutations in exon 2 counted by 50k reads/sample in deep tumor cells. (c) Counts of MED12 point mutations in 23 primary culture samples of myometrial cells without treatment (11 for acute (A) and 12 chronic (C) treatment controls). (d) Counts of MED12 point mutations in 24 primary culture samples of myometrial cells treated with PQ (100-200 μM) for up to 48 or 72 hrs (acute treatment). (e) Counts of MED12 point mutations in 15 primary culture samples of myometrial cells treated with PQ (100 μM) for 5 times, with each treatment cycle maintained for up to 48 hrs (chronic treatment). f Heatmap illustrated the relative frequency (high in red, low in blue) of MED12 exon2 mutations after acute (top) and chronic (bottom) PQ treatment. Mutation distribution of TCGA in each is marked on top. Red arrows highlight frequent mutations at c.127 and c.130-131