No evidence for transcriptional silencing of PTC-containing Ig-μ allele. (A) Schematic illustration of the pre-mRNA from the productively and nonproductively rearranged Ig-μ alleles as described in Figure 2A. (B) Analysis of the allele specificity of the two pre-mRNA TaqMan assays. Both assays were tested by qPCR as described in Figure 2B, and the amplicons were analyzed on a 2% agarose gel. (C) Relative pre-mRNA levels of productive and nonproductive allele were measured by RT-qPCR. Average values and standard deviations of four independent experiments with one run each are shown. For accurate calculation of the relative pre-mRNA abundances, the amplification rates were determined first. (D) Two independent pol II ChIPs were performed and the extracted DNA analyzed by qPCR using the two allele-specific TaqMan assays. The relative amount of DNA in the immunoprecipitated fraction was normalized to the input (input set to 1). (E) Two independent acetyl-H3 ChIPs were performed and analyzed as in D.
