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. 2016 Aug 10;7:12376. doi: 10.1038/ncomms12376

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Copyright © 2016, The Author(s)

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(a) Microarray analysis of pre-B cells from control and Vav-Cre Mef2c^fl/fl mice (9-months old) revealed 1,884 significantly (|FC|≥2 and P≤0.05) downregulated and 436 upregulated genes in the absence of Mef2c (n=3). (b) Cell cycle, DNA repair and transcription were among the most significantly enriched GO categories of genes downregulated in Mef2c-deficient pre-B cells. (c,d) Microarray and Q-PCR analysis of selected B-cell regulator that are significantly downregulated in Mef2c-deficient pre-B cells. (e) Microarray analysis of DNA DSB repair genes that are significantly downregulated in Mef2c-deficient pre-B cells are shown. (f) Q-PCR of key genes encoding DNA repair machinery validated defective expression in Mef2c-deficient pre-B cells. (d,f) n≥5 mice, data shown are the mean±s.d. of two or more independent experiments. **P<0.01, ***P<0.001, unpaired t-test. (g) Representative figures and quantification of alkaline comets revealed increased DNA damage in Mef2c-deficient pre-B cells (7–10-month old). n≥6 mice, data shown are the mean±s.e.m. of three independent experiments. ***P<0.001, unpaired t-test. About 728 control pre-B cells and 450 Mef2c-deficient pre-B cells were analysed. Scale bar, 20 μm. (h) Representative IF analysis of γH2AX in pre-B cells (7–10-months old) revealed that MEF2C is required for proper DSB repair in pre-B cells. n≥6 mice, data shown are the mean±s.d. of three independent experiments. Scale bar, 1 μm. ***P<0.001, unpaired t-test. (i) Quantification of alkaline comets revealed increased DNA damage in pro-B cells, but not CLP, sIgM⁺ bone marrow B cells, thymic T progenitors (DN1 and 2) or bone marrow myeloid cells in Mef2c-deficient mice (7–10-months old). n≥5 mice and data shown are the mean±s.e.m. of two or more independent experiments. More than 350 cells for each population were analysed. NS, not significant, **P<0.01, ***P<0.001, unpaired t-test. Both male and female mice were included for all experiments.