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