Fig. 6

Differential gene expression analysis of hMEC subpopulations treated with IR. a Experimental design to determine genes differentially expressed between CD10⁺ and MUC1⁺ enriched cultures (matched per donor) treated with IR. Freshly dissociated hMECs were cultured in vitro for 7 days (P1), then harvested and sorted based on expression of CD10 and MUC1 using FACS. Sorted cells were cultured for 5 days (P2), treated with 0, 2 and 5 Gy IR and then collected 2 and 24 hours post IR. RNA from all 12 samples was extracted and subjected to microarray analysis. The number of differentially expressed genes between the samples is indicated. b Principal component analysis of the expression of 19,472 filtered probes. Control (0 Gy) and 5 Gy treated samples of donor-matched (n = 5) CD10 and MUC1 sorted cultures are plotted. For clarity, donors are annotated consecutively: Q626, 1; Q695, 2; Q706, 3; Q827, 4; Q835, 5. c Subpopulation frequencies within hMEC cultures from MUC1⁺-sorted and CD10⁺-sorted cells performed in parallel to gene expression profiling. Mean subpopulation frequencies were calculated as the average percentage of total cell number per technical replicate for each sample (± SEM). DP frequencies of Q706 in CD10-sorted cells and MUC1-sorted cell K19⁺ frequencies were statistically different from all other donors (p <0.0001). d Dot plot demonstrating the Euclidian distance measured between 0 and 5 Gy matched samples at 24 hours of MUC1-sorted and CD10-sorted cultures. e Proportional Venn diagram depicting the number of shared and exclusive differentially expressed genes between MUC1 and CD10 samples 24 hours post 5 Gy IR treatment. f Normalised probe intensity from the gene expression analysis comparing CD10 and MUC1 samples at 24 hours after 5 Gy or no (0 Gy) IR treatment shown as the mean of five donors. g Relative expression of APOBEC3C in MUC1 and CD10 cultures before and after IR treatment shown as fold-change. The ratio of 5 Gy:0 Gy was significantly >1 for MUC1 (p = 0.034), whereas for CD10 the ratio was not significantly different from 1 (p = 0.84). FACS fluorescence-activated cell sorting, P1 passage 1, P2 passage 2