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. 2024 Oct 28;9:32. doi: 10.1038/s41536-024-00376-w

Fig. 4. Genomic imprinting of Mest persists in the blastema.

Fig. 4

A Schematic of the Mest genomic locus with exons as black boxes with exon number labeled in white; ΔEx3 denotes Mest-KO deletion. DMR (differentially methylated region) (gray circles); CTCF element (white box); miR335 (microRNA-335) (dark gray box); black bars denote regions of bisulfite sequencing. Location of Mest qPCR primers is shown with black arrows. B microRNA-335-5p qPCR analysis in Mest-WT, mKO, pKO, and KO 12 dpa blastemas. C Mest qPCR in Mest-WT, mKO, pKO, and KO 12 dpa blastemas. D–K Dot plots of bisulfite sequencing observed:predicted cytosine ratios for CTCF, ExonA, and Exon1 PCR amplicons shown in (A); each dot represents ratio for one sequenced clone. A ratio of 1 is fully methylated and a ratio of 0 is not methylated. D, F, H, J Bisulfite sequencing clones from embryonic e12.5 hindlimb Mest-WT and pKO DNA. E, G, I, K Bisulfite sequencing clones from 12 dpa blastemas of Mest-WT, mKO, pKO and KO mice. Error bars depict standard deviation; (*) p < 0.05, (**) p < 0.01, (***) p < 0.001, (****) p < 0.0001, ns not significant.