sheep polyclonal fibronectin antibody  (R&D Systems)

Journal: Science Advances

Article Title: ARID1A loss in neuroblastoma promotes the adrenergic-to-mesenchymal transition by regulating enhancer-mediated gene expression

doi: 10.1126/sciadv.aaz3440

Figure Lengend Snippet: ( A ) H3K27ac and BRG1 ChIP-seq tracks and RNA-seq tracks at PHOX2B and FN1 loci in control (red), ARID1A −/− #1 (blue), and ARID1A −/− #2 (black) NGP cells. ( B ) Western blot analysis of ARID1A, an adrenergic maker (PHOX2B), and three mesenchymal markers (FN1, SNAI2, and VIM) in control (Ctrl), ARID1A −/− #1, and ARID1A −/− #2 NGP cells. α-Tubulin was used as a loading control. ( C ) mRNA expression (RPKM of RNA-seq) of core regulatory circuitries of transcription factors specific for adrenergic (ADRN core TFs) or mesenchymal (MES core TFs) cells in control (Ctrl), ARID1A −/− #1, and ARID1A −/− #2 NGP cells. ( D ) GSEA to determine the enrichment of a generic gene signature for mesenchymal tumor in ARID1A mutant NGP cells. Genes are ranked by score and plotted along the x axis as vertical black bars. NES, normalized enrichment score; KO, knockout. ( E ) Cell viability [% relative to N , N ′-dimethylformamide (DMF)–treated cells, y value] of control (red), ARID1A −/− #1 (blue), and ARID1A −/− #2 (black). NGP cells were measured after treatment with various concentrations of cisplatin (0 to 20,000 nM, x value) for 72 hours, and half-maximal inhibitory concentration (IC 50 ) values are indicated. Values are means ± SD of triplicate experiments.

Article Snippet: Other antibodies and their sources were anti–α-tubulin mouse monoclonal (B-5-1-2) antibody (Sigma-Aldrich, #T6074), anti-PHOX2B mouse monoclonal (C-3) antibody (Santa Cruz Biotechnology, #SC-376993), anti-SNAI2 rabbit monoclonal (C19G7) antibody and anti-VIM rabbit monoclonal (D21H3) antibody (Cell Signaling Technology, #9585S and #5741S), and anti-FN1 sheep polyclonal antibody (R&D Systems, #AF1918).

Techniques: ChIP-sequencing, RNA Sequencing, Control, Western Blot, Expressing, Mutagenesis, Knock-Out, Concentration Assay