Escape from XCI in TLR7, RPS6KA3, CYBB, BTK, and IL13RA1 in Human pDCs
(A) Representative escape plots of female individuals heterozygous for the respective SNPs with each dot representing one pDC. All cells in one plot were derived from one female individual at the same time point. Blue dots are cells with monoallelic expression of the major allele, orange dots are cells with monoallelic expression of the minor allele, and green dots represent cells with biallelic expression (major and minor allele) in the same cell, reflecting escape from XCI. Black circles represent cells without any detectable mRNA. Percentages were calculated using all cells with detectable mRNA. The expression pattern was determined using the following SNPs: TLR7, rs3853839; RPS6KA3, rs7051161; CYBB, rs5964151; BTK, rs700; and IL13RA1, rs2495636 (n = 1 per gene).
(B) Representative escape plots of female individuals homozygous for the respective SNPs to ensure specificity of the cDNA SNP typing. Black circles represent cells without any detectable mRNA. Percentages were calculated using all cells with detectable mRNA. The expression patterns were determined using the following SNPs: TLR7, rs3853839; RPS6KA3, rs7051161; CYBB, rs5964151; BTK, rs700; and IL13RA1, rs2495636 (n = 1 per gene).
(C) Distribution of pDCs with biallelic expression for the five investigated genes across five females. The black bar represents the median percentage of pDCs with biallelic expression. Percentages were calculated using all cells with detectable mRNA. Dots with the same color represent the same female individual. F, female. For each gene and female, the percentage of biallelic expressing pDCs are as follows: TLR7: F2: 19%, F3: 37.3%, F4: 27.6%; RPS6KA3: F3: 22.9%, F4: 20.6%, F5: 17.9%; CYBB: F1: 15.4%, F3: 18.5%; BTK: F1: 22.2%, F2: 11.1%, F3: 17.2%, F4: 31.3%, F5: 5.6%; and IL13RA1: F1: 22.2%, F5: 30%.