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Fig. 4 | Genome Medicine

Fig. 4

From: Nuclear Wiskott–Aldrich syndrome protein co-regulates T cell factor 1-mediated transcription in T cells

Fig. 4

WASp functional status affects both basal and gene-specific transcription. a FUrd incorporation assays data for basal RNA polymerase II nascent transcripts in WT, WASp KO, and WASpI296T thymocytes as determined by intensity of FUrd-rich foci (red). The cell nucleus is labeled with DAPI (blue) and histone H3 (green). Note that mRNA transcripts can be seen in the cytoplasm already after 15 min. b Statistical analysis of FUrd assay results by nested ANOVA. Each dot represents one image. Top: WT n = 3, WASp KO n = 3 in one experiment; bottom: WT n = 4, XLNI296Tn = 4 in one experiment. c, d Gene expression profiling by RT-qPCR of WASp ChIP-seq genes (c) encoding TFs, other DNA- or RNA-binding proteins, and cytoskeletal network proteins performed on cDNA template of pooled samples: WASp KO n = 3 vs. WT n = 3 and (d) for common WASp-enriched genes in thymocytes and spleen CD4+ T cells. Expression in WASp KO thymocytes is shown and compared with expression level in WT thymocytes set to 1. Gene expression profiling by RT-qPCR was performed in three biological replicates: WASp KO n = 3 vs. WT n = 3 and in three technical replicates for each cDNA template sample with Ct variation between triplicates less than cutoff ± 0.25. Gene expression data: the mean of the technical triplicate, ΔCt, RQ value, and standard deviation between RQ of biological triplicates is indicated. See Additional file 1: Figure S5

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