Fig 2: Localisation of SMC1B mRNA with DAZL in human foetal ovary, by RNA in situ hybridisation in combination with immunohistochemistry.Columns show SMC1B (structural maintenance of chromosomes 1B) or DAZL (deleted in azoospermia-like), and merged images as indicated, with the right hand column showing higher power images at the indicated gestations and negative controls. Arrows identify somatic cell streams. Arrowheads identify germ-cell nests. Merge image; scale bar is 20 µm (the scale is the same for columns 1 and 2). Magnified image; scale bar is 10 µm.

Fig 3: 3'UTR-luciferase reporter assay. Increased luciferase activity is observed for target RNAs, but not an empty 3' untranslated region (3'UTR) or GAPDH 3'UTR, in the presence of DAZL (blue bar) relative to an empty vector control (magenta bar). No stimulation is observed when the R115G DAZL mutant is overexpressed (green bar). Firefly luciferase signals were normalised to Renilla luciferase signals. Mean ± SEM, n = 4, **P < 0.01 (Student's t-test).

Fig 4: Dazl is recruited to silenced transcription start sites in mES cells. (A) Annotation of Dazl ChIP-seq peaks, and the profile of Dazl peaks on the genome in comparison with Suz12, Aebp2 and H3K27me3 peaks in mES cells. The last ChIP profiles of Suz12, Aebp2 and H3K27me3 were obtained from (28). (B) Annotation of Dazl iCLIP peaks (top bar), and the intersect of Dazl ChIP-seq and iCLIP-seq peaks at the gene level (bottom Venn diagram). (C) Dazl peptides identified using SPACEmap procedure are shown by red (enriched in crosslinked fraction), blue (enriched in released fraction) and grey (statistically non-significant). (D) Proteins enriched by Dazl ChIP-SPACE in comparison to the IgG control were sorted by the abundance of the proteins (iBAQ). Histones and PRC2 components are shown by yellow and orange dots, respectively. Dazl and transcription factors are shown by red dots. (E) The volcano plot shows proteins identified by ChIP-SPACE and their sensitivity to RNase A treatment. Proteins that are affected by RNase treatment are named in the plot. (F) The schematic model of RBP interactions with chromatin based on Dazl data. Chromatin-associated RBPs form a condensed or confined zone probably via interactions among their IDRs with other components of chromatins. The RBPs that are in the periphery of the zone are sensitive to RNase treatment. The RBPs in the centre of the zone are resistant to RNase treatment. Transcription factors and other components of chromatin are probably recruited or trapped by the RBPs.