Journal: bioRxiv

Article Title: Multi-omics on truncating ASXL1 mutations in Bohring Opitz syndrome identify dysregulation of canonical and non-canonical Wnt signaling

doi: 10.1101/2022.12.15.520167

Figure Lengend Snippet: (A) Schematic representation of the ASXL1 transcript (ENST00000375687.10) and protein (GenBank: ASXL1; NM_015338.6 ; GRCh37), its functional domains, and mutations causing BOS. Mutations listed correspond to patients in this study and are tagged with a Pt identifier. (B) Peripheral blood and dermal fibroblasts were collected from BOS patients and controls. Dermal fibroblast samples underwent epigenomic assays for assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq), cleavage under targets & release using nuclease (CUT&RUN) and global transcriptome analysis using RNA sequencing (RNA-seq). Whole blood samples underwent DNA methylation analysis and global transcriptome analysis by RNA-seq. (C) Across the multi-omics assays and two specimen types we had varying degrees of overlap among assays. Six patients had one blood assay, five patients had both blood assays, four patients had multiple fibroblast assays, and three patients had data from assays across both specimen types. This totalled eighteen patients. (D) Western blot for representative BOS (n=3) and representative control (n=3) fibroblast whole cell lysate extracts shows no significant difference in total ASXL1 protein from ImageJ quantification. (E) Western blot for representative BOS (n=3) and representative control (n=3) fibroblast histone extracts shows no significant difference in H2AK119 ubiquitination, trimethylation of H3K4 and trimethylation of H3K27me3 from ImageJ quantification.

Article Snippet: For DNA purification, the DNA Purification Buffers and Spin Columns for ChIP and CUT&RUN (Cell Signaling, #14209), with elution in 50 ul.

Techniques: Functional Assay, Next-Generation Sequencing, RNA Sequencing Assay, DNA Methylation Assay, Whole Blood Assay, Western Blot

Journal: bioRxiv

Article Title: Multi-omics on truncating ASXL1 mutations in Bohring Opitz syndrome identify dysregulation of canonical and non-canonical Wnt signaling

doi: 10.1101/2022.12.15.520167

Figure Lengend Snippet: RNA-sequencing DESeq2 normalized counts show significant differential expression of PRXL2A between BOS and control samples in ( A ) blood (log2FC = 1.15, padj = 0) or ( B ) fibroblasts (log2FC = 1.96, padj = 0). ( C ) CUT&RUN identifies increased H3K4me3 binding at the PRXL2A transcriptional start site.

Article Snippet: For DNA purification, the DNA Purification Buffers and Spin Columns for ChIP and CUT&RUN (Cell Signaling, #14209), with elution in 50 ul.

Techniques: RNA Sequencing Assay, Expressing, Binding Assay

Journal: bioRxiv

Article Title: Multi-omics on truncating ASXL1 mutations in Bohring Opitz syndrome identify dysregulation of canonical and non-canonical Wnt signaling

doi: 10.1101/2022.12.15.520167

Figure Lengend Snippet: RNA-sequencing DESeq2 normalized counts show significant differential expression of GREM2 between BOS and control samples in ( A ) blood (log2FC = -2.48, padj = 0) or ( B ) fibroblasts (log2FC = -2.11, padj = 0). ( C ) CUT&RUN identifies increased H3K4me3 binding at the GREM2 transcriptional start site.

Article Snippet: For DNA purification, the DNA Purification Buffers and Spin Columns for ChIP and CUT&RUN (Cell Signaling, #14209), with elution in 50 ul.

Techniques: RNA Sequencing Assay, Expressing, Binding Assay