Figure 8.

Decreased H3K36me2 level in Miz1ΔPOZ correlates with sustained proliferation. A, Western blot analysis of H3K36me2 in sciatic-nerve homogenates from P30 control (Ctr) and P30 Miz1ΔPOZ mice and subsequent signal-intensity quantification in relation to Gapdh (right); n = 4 mice for each genotype. B, P30 Ctr and Miz1ΔPOZ animals were intraperitoneally injected seven times with EdU within 16 h for growth-fraction analysis (Fig. 4E). Confocal images were taken of longitudinal sciatic-nerve sections simultaneously stained for EdU (green) and H3K36me2 (red), maintaining the same image-acquisition settings. Dashed boxes highlight magnified regions. Arrowheads, EdU-stained nuclei. Scale bar, 50 μm. C, Corresponding staining-intensity quantification of confocal images representing the average H3K36^me2 intensity per area in relation to the average Hoechst signal of the same area for all nuclei (total) or only for EdU-positive nuclei (EdU⁺). n = 4 mice per genotype; analyzed nuclei: ≥1000 per sample. D, In vivo ChIP-qPCR analysis conducted on chromatin isolated from pooled sciatic nerves of 10 P30 Ctr or Miz1ΔPOZ mice using an H3K36me2 antibody and primers spanning a region close to the TSS or a coding region at the 3′ end of the indicated genes. For most genes and regions, an average fold enrichment compared with an unspecific IgG control >100, but at least 20 confirmed specificity in each experiment (data not shown). n = 3 independent experiments. A, C and D Asterisks indicate the results of a two-tailed, unpaired Student's t-test (*p < 0.05, **p < 0.01; ns = not significant).