Journal: Environmental Microbiology

Article Title: Evaluation of V2 18S rDNA barcode marker and assessment of sample collection and DNA extraction methods for metabarcoding of autotrophic euglenids

doi: 10.1111/1462-2920.15495

Figure Lengend Snippet: Average DNA concentrations measured spectrophotometrically (A) and fluorometrically (B). Symbols: DNE(Q) ‐ DNeasy Blood & Tissue (Qiagen); ENV(E) ‐ GeneMATRIX Environmental DNA & RNA Purification Kit (Eurx); SOI(E) ‐ GeneMATRIX Soil DNA Purification Kit (Eurx); SOI(O) ‐ E.Z.N.A. Soil DNA Kit (Omega Bio‐tek); WAT(O) ‐ E.Z.N.A. Water DNA Kit (Omega Bio‐tek); CTAB+D ‐ CTAB with DMSO; CTAB ‐ CTAB without DMSO; CHEL ‐ Chelex.

Article Snippet: Five commercially available DNA isolation kits were compared: DNeasy Blood & Tissue Kit (Qiagen; DNE(Q)), GeneMATRIX Environmental DNA & RNA Purification Kit (Eurx; ENV(E)), GeneMATRIX Soil DNA Purification Kit (Eurx; SOI(E)), E.Z.N.A.

Techniques: Purification, DNA Purification

Journal: Nature Communications

Article Title: Promotion of growth factor signaling as a critical function of β-catenin during HCC progression

doi: 10.1038/s41467-019-09780-z

Figure Lengend Snippet: β-Catenin does not promote early hepatocellular carcinoma (HCC) through Wnt signaling. a Immunoblot for β-catenin in triple knockout (TKO) HCC cells and Wnt3A-inducible mouse fibroblasts used as controls for non-active vs. active Wnt signaling (L cells; parental or Wnt3a+) treated with 25 μg/ml cycloheximide (CHX) or dimethyl sulfoxide (DMSO) for the time indicated (0–24 h). b Comparison of ctnnb1 complementary DNA (cDNA) sequence in control liver (CTRL) and TKO HCC. Regions highlighted in blue correspond to the phosphorylation sites. c Immunoprecipitation (IP) of immunoglobulin G (IgG) and Axin in TKO HCC cells. The presence of Axin, glycogen synthase kinase 3β (GSK3α/β), and phospho-β-catenin in the pull-down fraction was determined by immunoblot. d TKO HCC cells and as L cells (parental or Wnt3a+) were treated with 5 μM XAV939 for 48 h. The expression levels of Axin and β-catenin were determined by immunoblot. e Axin cDNA was overexpressed in TKO HCC cells and L cells (parental or Wnt3a+). The expression levels of Axin and β-catenin were determined by immunoblot. f Immunoblot for β-catenin and glutamine synthetase (GS) in TKO HCC cells treated with Wnt3A media. g Immunohistochemistry (IHC) for GS in control liver (CTRL) and TKO HCC. Black arrowheads indicate central vein (CV). The tumor zone in TKO HCC is delineated by a white dotted line. Scale bar = 50 μm. h Quantitative reverse transcription PCR (RT-qPCR) analysis for glul , tbx3 , lgr5 , and axin2 in control liver (CTRL; n = 4) and TKO HCC ( n = 9). i RT-qPCR analysis for glul , tbx3 , lgr5 , and axin2 in TKO HCC cells upon β-catenin knockdown (KD) with shβcat3-4 ( n > 3). j Immunoblot for β-catenin and GS in TKO HCC cells overexpressing dominant-negative of TCF4 (dnTCF4). k RT-qPCR analysis for glul , tbx3 , lgr5 , and axin2 in TKO HCC cells upon dnTCF4 overexpression ( n = 3). l Growth curve of TKO HCC cells upon dnTCF4 overexpression ( n = 2). m Fold difference in the number of TKO HCC organoids upon dnTCF4 overexpression ( n = 2). n Immunoblot of phospho β-catenin (Ser33/37/Thr41) and epithelial–mesenchymal transition (EMT) markers (Zeb1, Vimentin, and Claudin7) in control (CTRL) livers ( n = 3), primary TKO HCC tumors ( n = 4) and serially transplanted TKO HCC (Serial) tumors ( n = 4). Data are represented as mean ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001. n.s. not significant. See also Supplementary Fig.

Article Snippet: Human Flag-β-catenin pcDNA3, for human β-catenin overexpression, was a gift from Eric Fearon (Addgene plasmid #16828) . pWZL Blast DNE, for DNE overexpression, was obtained from Addgene (plasmid #18800) ; DNE sequence was further subcloned into MSCV-IRES-GFP (MigR1) vector.

Techniques: Western Blot, Triple Knockout, Comparison, Sequencing, Control, Phospho-proteomics, Immunoprecipitation, Expressing, Immunohistochemistry, Reverse Transcription, Quantitative RT-PCR, Knockdown, Dominant Negative Mutation, Over Expression