hand2  (Santa Cruz Biotechnology)

Journal: Cell Reports Medicine

Article Title: Histone lysine demethylase 4 family proteins maintain the transcriptional program and adrenergic cellular state of MYCN-amplified neuroblastoma

doi: 10.1016/j.xcrm.2024.101468

Figure Lengend Snippet: QC6352 impacts the chromatin accessibility of ADRN CRC TFs in MYCN -amplified cells (A and B) Adrenergic or mesenchymal median score in response to DMSO or 100 nM of QC6352 treatment for 48 h in adrenergic dominant cell lines (BE2C, SIMA, and KELLY) (A) and mesenchymal dominant cell lines (SK-N-AS and SK-N-SH) (B). KDM4 inhibition results in enhanced MES scoring with reduced ADRN signatures in all tested cell lines (red arrow indicates direction of signature change). (C) HOMER Motif analysis showing the predicted adrenergic CRC TF binding motifs in which genome-wide chromatin accessibility is impacted by QC6352 treatment of SIMA cells. (D) PHOX2A, ASCL1, and MYCN motif densities around ATAC-seq open chromatin regions (±1,000 bp) by categories in SIMA cells. Motif density was determined by the HOMER program and normalized to that in a background sequence of equal length. (E) STRING network analysis of transcriptional factors predicted to bind the regions with reduced chromatin accessibility by QC6352 treatment of SIMA cells. The expression threshold for genes encoding TFs is set as log2CPM > 1. CPM, counts per million mapped reads. Red color indicates ADRN CRC TFs. Blue color indicates MES CRC TFs. (F) IGV snapshots show chromatin accessibility at MYCN , HAND2 , PHOX2B , and ALK loci is reduced by QC6352 in SIMA and BE2C cells. (G) Density heatmaps for CUT&TAG of KDM4A-C genomic binding in BE2C cells, and ChIP-seq density heatmaps of the indicated histone marks in BE2C cells after 48 h treatment with control or 200 nM QC6352, ranked by MYCN read intensity within ± 5 kb of peak summits. MYCN ChIP-seq data were retrieved from GEO SuperSeries GSE94824. (H) The indicated epigenetic marks upregulated and downregulated by QC6352 treatment of BE2C cells for 48 h followed by ChIP-seq or CUT&TAG analyses. (I) The global distribution of H3K9me3, H3K36me3, H3K27Ac, H3K4me1, and H3K27me3 peaks at different regions of annotated genes. 3′ UTR, 3′ untranslated region; 5′ UTR, 5′ untranslated region; TSS, transcription start site. (J) Snapshot of using the IGV program displaying the KDM4s, H3K9me3, H3K36me3, H3K27me3, H3K27Ac, and H3K4me1 peaks at the PHOX2B and ASCL1 genomic loci.

Article Snippet: HAND2 (dHAND) , Santa Cruz , sc-398167, NA.

Techniques: Amplification, Inhibition, Binding Assay, Genome Wide, Sequencing, Expressing, ChIP-sequencing, Control

Journal: Cell Reports Medicine

Article Title: Histone lysine demethylase 4 family proteins maintain the transcriptional program and adrenergic cellular state of MYCN-amplified neuroblastoma

doi: 10.1016/j.xcrm.2024.101468

Figure Lengend Snippet:

Article Snippet: HAND2 (dHAND) , Santa Cruz , sc-398167, NA.

Techniques: Recombinant, Modification, Synthesized, Saline, Transfection, Apoptosis Assay, Purification, Transgenic Assay, Control, Expressing, Plasmid Preparation, Software

Journal: EMBO reports

Article Title: NONO enhances mRNA processing of super-enhancer-associated GATA2 and HAND2 genes in neuroblastoma.

doi: 10.15252/embr.202254977

Figure Lengend Snippet: Figure 6. NONO KD reduces mRNA and protein expression of GATA2 and HAND2, which may result from inappropriate processing and splicing in KELLY cells.

Article Snippet: Primary antibodies used were NONO (mouse monoclonal, in-house made), GATA2 (CG2-96) (mouse monoclonal, Santa Cruz, sc-267), HAND2 (A-12) (mouse monoclonal, Santa Cruz, sc-398167) and SFPQ (mouse monoclonal, Merck, P2860).

Techniques: Expressing

Journal: Nature communications

Article Title: Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma.

doi: 10.1038/s41467-022-29311-7

Figure Lengend Snippet: Fig. 2 scRNA-sequencing of early LPM reveals a heterogeneous progenitor pool. A LPM-marking drl:mCherry-positive cells of zebrafish embryos at tailbud stage, FACS-isolated, and sequenced using CEL-Seq2. B SPIM projection of drl:mCherry embryo at tailbud, labeling the LPM. C UMAP plot displaying 15 LPM cell clusters, colored by subpopulation: cardiopharyngeal (1,2), heart field, anterior vasculature, hemangioblasts/kidney (1,2), head mesoderm, hatching gland, hand2-high (1–4), and endoderm (1–3) as distinct group. D Approximate anterior-to-posterior orientation based on cdx4 (RNA ISH at 5 ss). E Dotplot including key cell fate markers to annotate clusters. Dots colored by column-scaled mean expression (log-transformed library-size-normalized counts) and sized by expression frequency (fraction of cells with non-zero counts); rows and clusters ordered by hierarchical clustering of scaled expression values. F UMAP plots of several genes co-expressed with hand2 or among cluster-determining genes in four hand2-high clusters. Cells colored by scaled expression values using lower/upper 1%-quantile boundaries. G–K Whole-mount gene expression analysis of select transcripts enriched in hand2-high cells by fluorescent in situ hybridization (ISH) (G, H) and colorimetric mRNA ISH (I–K). Fluorescent ISH of meis3 (G), foxh1 (H) with hand2 at 4 ss, revealing overlap in posterior LPM (magnified regions from dashed boxes). meis3 ISH at 10 ss (I), gata5 at 12 ss (J), and sfrp5 at 18 ss (K) showing expression in lateral-most LPM sprawling outwards (arrowheads). Scale bar G, H 100 μm, I–K 250 μm.

Article Snippet: The sections were blocked by incubating with 2% BSA 1× PBS supplemented with 1% horse serum at room temperature and incubated with HAND2 antibody (Santa Cruz Biotechnology A-12, Sc-398167, at dilution 1:100) overnight at 4 °C.

Techniques: Sequencing, Isolation, Labeling, Expressing, Transformation Assay, Gene Expression, In Situ Hybridization

Journal: Nature communications

Article Title: Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma.

doi: 10.1038/s41467-022-29311-7

Figure Lengend Snippet: Fig. 4 Primordial germ cells migrate with, and home within, the hand2-positive LPM. A UMAP plots of cxcl12a, hand2, and pax2a. Cells are colored by scaled expression values using lower and upper 1%-quantiles as boundaries. B, C Snapshots of time-lapse movie (Supplementary Movie 1) showing drl:EGFP with primordial germ cell (PGC) marker mCherry-f′-nos3′UTR, with 3D renderings at 60% epiboly, 1 ss, and 10 ss (B) and maximum intensity Mercator projections for the same time points (C). The PGCs migrate from four different clusters during gastrulation into two bilateral clusters during somitogenesis (boxed regions). Note the sparse (lost) PGCs located within the endoderm (asterisks). Animal side is for 60% epiboly to the top, anterior for 1 ss and 10 ss (B) to the top and (C) to the left. D Imaging of hand2:EGFP with mCherry-f′-nos3′UTR showing a dorsal view at 24 hpf: the PGC clusters fall completely within the hand2:EGFP domain. The outlines of the yolk extension are highlighted. E Segmented 3D rendering of a hand2:EGFP; mCherry-f′-nos3′ UTR double-positive embryo stained with anti-Pax2a (yellow) and DAPI (blue) at 28 hpf. F A schematic transverse section of a 24 hpf embryo showing two bilateral PGC clusters (magenta) lateral of the developing gut, ventral-lateral of the Pax2a-positive developing pronephros (yellow) within the cxcl12a-high hand2-positive LPM/coelomic epithelium (green). Scale bar D, E 250 μm.

Article Snippet: The sections were blocked by incubating with 2% BSA 1× PBS supplemented with 1% horse serum at room temperature and incubated with HAND2 antibody (Santa Cruz Biotechnology A-12, Sc-398167, at dilution 1:100) overnight at 4 °C.

Techniques: Expressing, Marker, Imaging, Staining