Fig. 2

TLR7 and TLR8 evade X chromosome inactivation in the monocytes and CD4⁺ T cells of women. A–D RNA FISH analysis of CD14⁺ monocytes from female donors. The images show confocal microscopy planes of cell nuclei after hybridization with fluorescent probes for transcripts arising from TLR7 (green), TLR8 (red), and the marker gene triad transcribed from the Xa only (PGK1, CFP, MSN; pink). Nuclei are counterstained with 4ʹ,6-diamidino-2-phenylindole (DAPI; blue). The arrow heads in A, C indicate transcript foci from the two alleles of TLR7 or TLR8. In B, D, the two genes are transcribed only from the Xi (arrowheads). The TLR7 or TLR8 hybridization pattern is schematized to the right of each row. E–H Quantification of the escape from XCI for TLR7 and TLR8 from monocytes (E, F) and CD4⁺ T cells (G, H). The forest plots show the percentage of XCI escape in monocytes or T cells and its 95% confidence interval (CI) in individual female donors (n = 6); the red diamond denotes the meta-analytical group mean and 95% CI. I, J Confocal microscopy planes of cell nuclei after RNA FISH with TLR7 (I, pink), TLR8 (J, red), and XIST (green, painting the Xi) probes. Nuclei are counterstained with DAPI (blue). The TLR7 and TLR8 transcriptional foci adjacent to the Xi territory confirm that both genes evade XCI