Journal: Journal of Developmental Biology

Article Title: Asxl2 −/− Mice Exhibit De Novo Cardiomyocyte Production during Adulthood

doi: 10.3390/jdb4040032

Figure Lengend Snippet: The Asxl2 −/− hearts exhibit elevated proliferative activity in non-cardiomyocytes. ( A ) Representative image showing EdU, wheat germ agglutinin (WGA), and 4′,6-diamidino-2-phenylindole (DAPI); ( B ) Quantification of BrdU + nuclei in left ventricle (LV) at 8-weeks (wildtype: n = 3, 3 non-consecutive sections/heart, 116 BrdU + , 30,045 total nuclei; Asxl2 −/− : n = 3, 3 non-consecutive sections/heart, 170 BrdU + , 36,564 total nuclei) and 12-weeks (wildtype: n = 3, three non-consecutive sections/heart, 35 BrdU + , 13,853 total nuclei; Asxl2 −/− : n = 3, three non-consecutive sections/heart, 127 BrdU + , 15,250 total nuclei); ( C ) Quantification of average number of EdU + per 20× image field at 12-weeks (wildtype: n = 3, three non-consecutive sections/heart, 75 images; Asxl2 −/− : n = 3, three non-consecutive sections/heart, 75 images); ( D ) Representative image of EdU, vimentin and DAPI labeling; ( E , E′ ) Representative image of EdU, cTnT and DAPI labeling; ( F ) Quantification of Ki67 + nuclei in LV at 12-weeks (wildtype: n = 3, three non-consecutive sections/heart, 51 Ki67 + , 67,727 total nuclei; Asxl2 −/− : n = 3, three non-consecutive sections/heart, 236 Ki67 + , 62,124 total nuclei); ( G – G″ ) Representative image of Ki67, WGA and cTnT labeling. Arrows indicate proliferative cell nuclei. Bars indicate standard deviation. * p -Value (Student’s t -test) < 0.05.

Article Snippet: Primary antibodies used on paraffin sections include rat-α-BrdU (1:100, Abcam, Cambridge, MA, USA, ab6326), mouse-α-cTnT (1:100, Thermo Scientific, MS-295), rabbit-α-Ki67 (1:400, Abcam, ab15580), rabbit-α-vimentin (1:100, Abcam, ab92547), rabbit-α-CD31 (1:40, Lifespan BioSciences, Seattle, WA, USA, LS-B1932), rabbit-α-α-smooth muscle actin (1:800, Sigma, A2547), rabbit-α-NKX2-5 (1:100, Abcam, ab22611), rabbit-α-MEF2C (1:100, Abcam, ab64644), rabbit-α-GATA4 (1:100, Santa Cruz, Dallas, TX, USA, sc-9053), and rabbit-α-connexin43 (1:200, Santa Cruz, sc-6560).

Techniques: Activity Assay, Labeling, Standard Deviation

Journal: Journal of Developmental Biology

Article Title: Asxl2 −/− Mice Exhibit De Novo Cardiomyocyte Production during Adulthood

doi: 10.3390/jdb4040032

Figure Lengend Snippet: EdU-labeled cells give rise to cardiomyocytes in Asxl2 −/− hearts after 4-week chase. ( A ) Schematic of the lineage tracing assay used to assess the differentiation potential of the proliferating non-cardiomyocytes observed at 12 weeks of age; ( B ) Quantification of the percentage of EdU + cells that are cTnT − NKX2-5 + and cTnT + NKX2-5 + in the left ventricle following the chase; ( C ) Representative image of a mononuclear, EdU + cTnT + NKX2-5 + cell in a longitudinal orientation; ( C′ ) The same cell in c, shown in a z-stack image; ( C′′ ) Cross-sections of the z-stack image in c′; ( D – D′′ ) Representative image of an EdU + cTnT + cell, delineated with wheat germ agglutinin (WGA), in a transverse heart section; ( E – E′′ ) Representative image of EdU, WGA and labeling for the gap junction protein, connexin 43 (CX43). The arrow indicates the CX43 + membrane between two cardiomyocytes. Sample size: n = 3 animals per genotype; three non-consecutive sections/heart; analyzed LV from whole section-stitched images; 617 and 755 EdU + cells were examined on wildtype and Asxl2 −/− heart sections, respectively. Bars represent standard deviation. * p- Value (Student’s t -test) < 0.05.

Article Snippet: Primary antibodies used on paraffin sections include rat-α-BrdU (1:100, Abcam, Cambridge, MA, USA, ab6326), mouse-α-cTnT (1:100, Thermo Scientific, MS-295), rabbit-α-Ki67 (1:400, Abcam, ab15580), rabbit-α-vimentin (1:100, Abcam, ab92547), rabbit-α-CD31 (1:40, Lifespan BioSciences, Seattle, WA, USA, LS-B1932), rabbit-α-α-smooth muscle actin (1:800, Sigma, A2547), rabbit-α-NKX2-5 (1:100, Abcam, ab22611), rabbit-α-MEF2C (1:100, Abcam, ab64644), rabbit-α-GATA4 (1:100, Santa Cruz, Dallas, TX, USA, sc-9053), and rabbit-α-connexin43 (1:200, Santa Cruz, sc-6560).

Techniques: Labeling, Membrane, Standard Deviation