Fig. 4. MED12 Gly-44 mutations lead to abnormal DNA repair activity and render cells sensitive to DNA-damaging chemotherapeutic agents.

a Flow cytometry profiles show a representative image of triplicate data showing the rate of EdU incorporation and DNA content analysis in WT and MED12 mutant cells. b Western blots show γH2AX and β−Actin protein levels in WT and mutant cells. Whole membrane blots are shown in Supplementary Fig. 12. c The images (quantified in dot plots) show relative immunofluorescence of γH2AX signal intensity in control and Carboplatin-treated WT and MED12 mutant cells. d Immunohistochemistry images show γH2AX staining in normal and MED12 mutant fibroid tumor tissue microarray. The staining intensity in individual cells was assessed by machine learning-assisted segmentation and quantification (see methods). e, f The dot plots show overall (e) and smooth muscle-specific (f) γH2AX IHC signal intensity across ten distinct UFs tissue specimens. g The Incucyte live-cell imaging results show relative rates of apoptosis (The Incucyte® Caspase-3/7 dye) WT and MED12 mutant cells measured over more than 3 days in control and Carboplatin-rated cells. For statistical analysis in a, c, e, f, a two-sided unpaired t test has been used. Error bars indicate the standard error of the mean.