Journal: BMC Nephrology

Article Title: Fine needle aspirates of kidneys: a promising tool for RNA sequencing in native and transplanted kidneys

doi: 10.1186/s12882-018-1012-4

Figure Lengend Snippet: RNA yield and RNA quality of FNA samples (G19-G25) and of corresponding core biopsy samples (G16). a . RNA yield (ng/sample) was higher in G16 biopsies compared to all FNA (G19–25) samples (***p = 0,0003; ****p < 0,0001) . b . RNA quality by RIN values, and c . RNA quality by DV200 values

Article Snippet: Also, the total RNA yield from some of these G25 size aspirations was, however, at the threshold of 50 ng/sample required for RNA input for both sequencing panels according to Illumina ( http://www.support.illumina.com/content/illumina-marketing/en/products/truseq-targeted-rna-expressionwnt-panel.html and http://www.support.illumina.com/content/illumina-marketing/en/products/truseq-targeted-rna-expression-cell-cycle-panel.html ).

Techniques:

Journal: Oncogene

Article Title: Temporal activation of WNT/β-catenin signaling is sufficient to inhibit SOX10 expression and block melanoma growth

doi: 10.1038/s41388-020-1267-7

Figure Lengend Snippet: a Representative western blots of the indicated proteins in the MAPK inhibitor resistant human melanoma cell culture M12, at different timepoints of CHIR99021 (6 μM) treatment (timepoints are shown directly in the figure). GAPDH was used as loading control ( n = 3). b RNA levels in MAPK inhibitor sensitive (M98) human melanoma cell culture (ΔCt to GAPDH expression) after 4, 8, and 12 h of CHIR99021 (6 µM) treatment are shown compared with DMSO (ctrl) ( n = 3). c Representative western blot of the indicated proteins in the MAPK inhibitor sensitive human melanoma cell culture (M98). A representative western blot for the indicated proteins of M98 human melanoma cell culture stably expressing shSOX10 or shMITF constructs cultured in presence or absence of CHIR99021 for 24 h, 6 μM. Nssh represents the scrambled shRNA used as control construct ( n = 3). d Representative western blot of the indicated proteins in presence of CHIR99021 (6 μM) and/or MG-132 (20 μM) (in single or double treatments) for 16 h. GAPDH was used as a loading control ( n = 3). e Heatmap demonstrating the gene expression of SOX10 and Wnt target genes of M98 and M00 (MAPK inhibitor sensitive cell cultures) and M11 (resistant to MAPK inhibitor) in either presence or absence of CHIR99021. Data on the heat map represent log2 of sequencing reads assigned to genes. Statistical significance was determined by unpaired, two-tailed Student’s t test in b . * P < 0.05, ** P < 0.01, *** P < 0.001. In each panel, n indicates the number of independent experiments performed.

Article Snippet: Annotated WNT and SOX10 transcriptional target gene lists were obtained from Metacore ( https://portal.genego.com/ ).

Techniques: Western Blot, Cell Culture, Control, Expressing, Stable Transfection, Construct, shRNA, Gene Expression, Sequencing, Two Tailed Test

Journal: Oncogene

Article Title: Temporal activation of WNT/β-catenin signaling is sufficient to inhibit SOX10 expression and block melanoma growth

doi: 10.1038/s41388-020-1267-7

Figure Lengend Snippet: CHIR99021 is a small chemical compound that competes with ATP for the binding site in GSK3α/β kinase and thereby inhibits GSK3α/β, leading to the activation of WNT signaling. SOX10 protein levels are reduced following pharmacologic, acute β-catenin stabilization, and this is mediated via proteasomal degradation.

Article Snippet: Annotated WNT and SOX10 transcriptional target gene lists were obtained from Metacore ( https://portal.genego.com/ ).

Techniques: Binding Assay, Activation Assay