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. 2015 May 22;10(5):e0127839. doi: 10.1371/journal.pone.0127839

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© 2015 Stein et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Fig 8 — Immunoblot analysis for PTIP and H3K4me3 was performed to determine the impact of PTIP deletion and TAC on global H3K4me3 levels and PTIP expression. As shown in panel A, PTIP deletion results in a significant attenuation in global H3K4me3 marks when normalized to total histone H3 levels. TAC had no significant impact on PTIP levels or H3K4me3 levels. To determine whether PTIP regulates H3K4me3 levels at the promoter region of specific genes, ChIP-qPCR was performed to assess H3K4me3 enrichment at the promoter region of ADRA1A and SCN4B in PTIP- and PTIP+ mice (panel B). ChIP was performed with an anti-H3K4me3 antibody (black) and a non-specific Rabbit IgG antibody (white). Data reveal a significant decrease enrichment of H3K4me3 marks in PTIP- hearts (n = 6) as compared to PTIP+ hearts (n = 6) at the Adra1a and SCN4b promoter. The amplified ChIP-qPCR products were run on a gel with a 2% input (panel B, below). Data shown are means ± SEM.