Journal: Haematologica

Article Title: Transforming activities of the NUP98-KMT2A fusion gene associated with myelodysplasia and acute myeloid leukemia

doi: 10.3324/haematol.2019.219188

Figure Lengend Snippet: iNUP98-KMT2A expression impairs cell cycle progression of murine embryonic fibroblasts and bone marrow-derived hematopoietic stem and progenitor cells. (A) Murine embryonic fibroblasts (MEF), derived from iNUP98-KMT2A and wildtype (WT) control (CTRL) littermate mice were cultured in vitro in the presence of doxycycline (DOX) (1 μg/mL). iNUP98-KMT2A expression is shown relative to the level of GAPDH expression. * P <0.05, unpaired t -test, n=3. (B) Flow cytometry-based cell cycle analysis showed increased G 1 and decreased G 2 /M fractions of in vitro -cultured iNUP98-KMT2A+ MEF compared to WT controls. * P <0.05, unpaired t -test, n=3. (C) iNUP98-KMT2A and WT MEF cultured for eight passages in the presence of DOX (1 μg/mL) were stained for senescence-associated B-galactosidase activity with X-Gal (left panel). The number of X-Gal + cells in the culture was quantified (right panel). Images and counts are representative of three biological replicates. Scale bars: 100 μm. ** P <0.01, unpaired t-test, n=3. (D) Differential mRNA expression from early (passages 1-2) and late (passages 8-10) passaged WT and iNUP98-KMT2A MEF analyzed by a RT2 PCR array. Significant ( P <0.05) changes are highlighted in red. (E) Validation of differentially expressed genes in MEF by quantitative polymerase chain reaction analysis in WT and iNUP98-KMT2A hematopoietic stem and progenitor cells after exposure to DOX (1 μg/mL) in vitro for 48 h. *<0.05, unpaired t -test, n=3.

Article Snippet: Using fluorescent in situ hybridization and reverse transcription quantitative polymerase chain reaction (PCR), Kaltenbach et al . found that inv(11)(p15q23) leads to fusion of the NUP98-FG -repeats to almost the entire KMT2A open reading frame (ORF).

Techniques: Expressing, Derivative Assay, Control, Cell Culture, In Vitro, Flow Cytometry, Cell Cycle Assay, Staining, Activity Assay, Biomarker Discovery, Real-time Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Hearts of Dystonia musculorum Mice Display Normal Morphological and Histological Features but Show Signs of Cardiac Stress

doi: 10.1371/journal.pone.0009465

Figure Lengend Snippet: Primers used for QPCR experiments.

Article Snippet: Quantitative polymerase chain reaction (QPCR) (MJ Research) was performed in triplicate for each sample using primer pairs as detailed in .

Techniques:

Journal: PLoS ONE

Article Title: Hearts of Dystonia musculorum Mice Display Normal Morphological and Histological Features but Show Signs of Cardiac Stress

doi: 10.1371/journal.pone.0009465

Figure Lengend Snippet: mRNA was isolated from wt and dt hearts and QPCR was performed to assess transcript levels of other plakin proteins expressed in cardiac tissue.

Article Snippet: Quantitative polymerase chain reaction (QPCR) (MJ Research) was performed in triplicate for each sample using primer pairs as detailed in .

Techniques: Isolation

Journal: International Journal of Molecular Medicine

Article Title: BMS-345541 inhibits airway inflammation and epithelial-mesenchymal transition in airway remodeling of asthmatic mice

doi: 10.3892/ijmm.2018.3762

Figure Lengend Snippet: BMS-345541 inhibits epithelial-mesenchymal transition in asthma. mRNA levels of E-cadherin and vimentin in mouse lung tissues were measured by (A) reverse transcription-quantitative polymerase chain reaction analysis. (B) Protein levels were measured by western blot analysis for (C) E-cadherin and (D) vimentin. Data are presented as the mean ± standard deviation (n=8 per group). # P<0.01 and ## P<0.001, OVA group compared with the control group; * P<0.05 and ** P<0.01, OVA + BMS-345541 group compared with the OVA group. OVA, ovalbumin; DMSO, dimethyl sulfoxide.

Article Snippet: The reagents used were as follows: OVA (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany), acetylcholine chloride (Ach; Sigma-Aldrich; Merck KGaA), phosphate-buffered saline (PBS; Sigma-Aldrich; Merck KGaA), sodium pentobarbital (Sinopharm Chemical Reagent Co., Ltd., Shanghai, China), BMS-345541 (Abcam, Cambridge, MA, USA), E-cadherin antibody (cat. no. ab76055; Abcam), vimentin antibody (cat. no. ab92547, Abcam), β-actin antibody (cat. no. ab179467; Abcam), glyceraldehyde 3-phosphate dehydrogenase (GADPH; Sigma-Aldrich; Merck KGaA), dimethyl sulfoxide (DMSO; Sigma-Aldrich; Merck KGaA), diaminobenzidine (DAB; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), an enzyme-linked immunosorbent assay (ELISA) kit (Bio-Rad Laboratories, Inc., Hercules, CA, USA), Pierce Enhanced Chemiluminescence (ECL) western blot substrate (Thermo Fisher Scientific, Inc., Waltham, MA, USA), a reverse transcription kit (Promega Corporation, Madison, WI, USA), a quantitative polymerase chain reaction (qPCR) kit (TransStart Green; Beijing Transgen Biotech Co., Ltd., Beijing, China), a bicinchoninic acid (BCA) protein assay kit and radioimmunoprecipitation assay (RIPA) lysis buffer (Beijing ComWin Biotech Co., Ltd., Beijing, China).

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation, Control