Figure 3.

Cortex and spinal cord NSPCs feature differential deposition of H3K27me3, H3K4me3 and H3K9/14ac at the level of both Hoxb4–6 5' regions and RAREs. (a–f) Real-time PCR quantification of ChIP assays in cortex and spinal cord NSPCs using control (IgG), anti-H3K27me3 (αH3K27me3), anti-H3K4me3 (αH3K4me3) or anti-H3K9/14ac (αH3K9/14ac) antibodies. Primer pairs used for real-time PCR target either the genomic regions 5' of Hoxb4 (a), Hoxb5 (b), Hoxb6 (c), or ENE-RARE (d), B4U-RARE (e), DE-RARE (f), as indicated. A significant increase in H3K4me3 and H3K9/14ac levels and significantly lower H3K27me3 levels are detectable in spinal cord NSPCs relative to cortex NSPCs. No changes between spinal cord and cortex NSPCs are detectable following control ChIP (ChIP IgG). Results are shown as the mean of the log₂-transformed ratio between spinal cord and cortex NSPCs in three to six biological replicates, following normalization to a reference amplicon as described in the Material and methods section. Error bars show s.e.m. *p ≤ 0.05; **p < 0.01; ***p < 0.001; n.s., non-significant (p > 0.05) according to a two-tailed Student's t-test performed between spinal cord and cortex NSPC samples.