Figure 7.

Identification of Kdm8 and Znhit3 as direct Miz1 target genes in vitro and in vivo. A, In vitro ChIP-qPCR analysis conducted on chromatin isolated from the mSC80 Schwann cell line, using either a specific α-Miz1 antibody (H190 sc-22837) or an unspecific IgG control. Primers span either the TSS, including the Miz1 binding motif, or a randomly chosen upstream region (+1000 bp). n = 4 independent experiments. B, RNAi-mediated Miz1 knockdown in mSC80 cells using siRNA directed against Miz1 (siMiz1) or unspecific control siRNA (siSCR). Efficient knockdown of endogenous Miz1 48 h after transfection was confirmed on Western blot using the 10E2 antibody. Gene expression of the Miz1 target genes Kdm8 and Znhit3 relative to Gapdh 48 h after siRNA transfection, determined by qRT-PCR analysis. n = 4 independent experiments. C, In vivo ChIP-qPCR analysis performed on chromatin isolated from pooled sciatic nerves of 10 P30 Ctr mice, applying the same antibodies and primers as in A. n = 3 independent experiments. D, Kdm8 and Znhit3 gene expression measured by qRT-PCR analysis in P30 Ctr and Miz1ΔPOZ sciatic nerves. n = 4 mice per genotype. A–D Asterisks indicate the results of a two-tailed, unpaired Student's t-test (*p < 0.05, ***p < 0.001).