Fig. 6
At class II promoters NM1 maintains and preserves active epigenetic marks by direct recruitment of the HAT PCAF and the HMT Set1/Ash2. a-d ChIP and qPCR analysis on chromatin isolated from NM1 knockdown MEFs (NM1 RNAi) and control MEFs (scrRNAi), using antibodies against PCAF, Set1/Ash2, H3K9Ac, H3K27Ac, H3K4me3 and H3K4me1 as well as non-specific IgGs. In all cases, qPCR analysis was performed with primers amplifying (a, c) the mouse Rpl19 gene promoter and (b, d) the mouse Rad9a gene promoter. All ChIP experiments were successfully performed at least three times (n = 3). The values are presented as the percentage of the input signal for each primer pair. Error bars represent standard deviations. Upon NM1 gene knockdown, significant changes were detected for PCAF, Set1/Ash2 and for the epigenetic marks H3K9Ac, H3K27ac and H3K4me3. In panel a, pPCAF = 0.00472 (**), pSet1/Ash2 = 0.0214 (*); in panel b, pPCAF = 0.01 (**), pSet1/Ash2 = 0.0026 (***); in panel c, pH3K9ac = 0.000353 (***), pH3K27ac = 5.93E-05 (****), pH3K4me3 = 0.000271 (***); in panel d, pH3K9ac = 8.04E-05 (****), pH3K27ac = 4.92E-05 (****), pH3K4me3 = 0.0046 (**). Significances were obtained by Student’s t-test, two-sample equal variance. ns = non-significant ChIP-Seq chromatin immunoprecipitation and deep sequencing, HAT histone acetyl transferase, HMT histone methyl transferase, MEFs mouse embryonic fibroblasts, NM1 nuclear myosin 1c, qPCR quantitative polymerase chain reaction