Journal: Scientific Reports

Article Title: Establishment of a stable transfection method in Babesia microti and identification of a novel bidirectional promoter of Babesia microti

doi: 10.1038/s41598-020-72489-3

Figure Lengend Snippet: Growth curve analysis. Pre-patent period analyses for the transgenic and wild type B. microti parasites. The growth of wild type and transgenic B. microti parasites are similar. Each data point represents an average parasitemia of five mice with SD. The p-value (p = 0.5) was calculated using t-test. A p-value of p > 0.05 represents statistically not significant.

Article Snippet: Briefly, 100 μl of B. microti (grey strain, ATCC 30221D) infected blood (~ 10% parasitemia) was collected from infected mice in a 1.5 ml tube with 10 μl of heparin (10,000 units/ml).

Techniques: Transgenic Assay

Journal: Scientific Reports

Article Title: Establishment of a stable transfection method in Babesia microti and identification of a novel bidirectional promoter of Babesia microti

doi: 10.1038/s41598-020-72489-3

Figure Lengend Snippet: Prepatent period for BM-WT and BM-GFP/mCherry parasites.

Article Snippet: Briefly, 100 μl of B. microti (grey strain, ATCC 30221D) infected blood (~ 10% parasitemia) was collected from infected mice in a 1.5 ml tube with 10 μl of heparin (10,000 units/ml).

Techniques:

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