Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Replication Stress Response Modifies Sarcomeric Cardiomyopathy Remodeling

doi: 10.1161/JAHA.121.021768

Figure Lengend Snippet: ( A ) Representative immunofluorescence staining of Ki67 (red) and γH2AX (green) and nuclei labeled with 4′,6‐diamidino‐2‐phenylindole (DAPI) (blue) in control (Ctl) and Mybpc3 −/− left ventricular (LV) tissue at postnatal day (P) 7. Bar=5 µm. ( B ) Percentage of Ki67‐γH2AX colocalized nuclei per total Ki67‐positive nuclei in Ctl (n=4) and Mybpc3 −/− (n=4) LV tissue. ( C ) Representative immunofluorescence staining of cardiomyocyte γH2AX in Ctl and Mybpc3 −/− LV tissue from mice administered the cyclin‐dependent kinase (CDK) 4/6 inhibitor (inhib.) PD‐0332991 (150 mg/kg per day). DNA damage marker γH2AX (red), cardiomyocyte (CM) marker pericentriolar material 1 (PCM1) (green), and nuclei marker DAPI (blue). Bar=100 µm. ( D ) Quantification of γH2AX CM nuclei in Ctl (n=5–6) and Mybpc3 −/− (n=5–6) LV tissue from untreated mice or from mice administered with CDK4/6 inhib. Minimum of 100 CM nuclei/sample. ( E ) Western blot of phosphorylated ataxia telangiectasia and rad3 related (ATR) (p‐Ser428 ATR) and total ATR in P7 Mybpc3 −/− myocardial tissue from untreated or CDK4/6 inhib. treated mice. ( F ) Relative quantification of p‐Ser428 ATR from untreated or CDK4/6 inhib. groups normalized to total ATR (n=5). ( G ) Measurement of 8‐hydroxy‐2'‐deoxyguanosine (8‐OHdG) levels (ng/µg of DNA) through competitive ELISA from P7 Ctl (n=4) and Mybpc3 −/− (n=4) myocardial tissue. All results are shown as mean±SEM.

Article Snippet: Genomic DNA was extracted from snap‐frozen P7 myocardial tissue, according to commercially available DNA extraction kit (Qiagen, 69504).

Techniques: Immunofluorescence, Staining, Labeling, Inhibition, Marker, Western Blot, Competitive ELISA

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Replication Stress Response Modifies Sarcomeric Cardiomyopathy Remodeling

doi: 10.1161/JAHA.121.021768

Figure Lengend Snippet: ( A ) Schematic of Mybpc3 −/− /p53 fl/fl Myh6Cre +/− double‐null murine model generated by crossing a p53 fl/fl Myh6Cre +/− mouse with an Mybpc3 −/− mouse. ( B ) Western blot of p53 from control (Ctl) (n=3), Mybpc3 −/− (n=3), and Mybpc3 −/− /p53 fl/fl Myh6Cre +/− (n=3) myocardial tissue at postnatal day (P) 25. Western blot of β‐actin used as loading control. ( C ) Measurement of p53 target gene expression (mouse double minute 2 [Mdm2], cyclin‐dependent kinase inhibitor 1 [Cdnk1a], growth differentiation factor 15 [Gdf15], and growth arrest and DNA damage inducible α [Gadd45a]) at P25 in Ctl (n=6), Mybpc3 −/− (n=6), and Mybpc3 −/− /p53 fl/fl Myh6Cre +/− (n=6) left ventricular (LV) tissue RNA. The genes of interest were normalized to Rpl32 expression. Fold changes are shown relative to Ctl gene expression. Echocardiography assessment of interventricular septal thickness at end diastole (IVSd) ( D ), LV posterior wall thickness at end diastole (LVPWd) ( E ), LV internal diameter at end diastole (LVIDd) ( F ), and fractional shortening (FS) ( G ) in Ctl (n=6), p53 fl/fl Myh6Cre +/− (n=4), Mybpc3 −/− (n=13), and Mybpc3 −/− /p53 fl/fl Myh6Cre +/− (n=13) mice at P25. Heart weight (HW) ( H ) and HW/tibia length (TL) ratio ( I ) from Ctl (n=7), p53 fl/fl Myh6Cre +/− (n=6), Mybpc3 −/− (n=7), and Mybpc3 −/− /p53 fl/fl Myh6Cre +/− (n=6) mice at P25. ( J ) Representative immunohistochemical staining using wheat‐germ agglutinin (green) and 4′,6‐diamidino‐2‐phenylindole (blue) of LV tissue from Ctl, Mybpc3 −/− , and Mybpc3 −/− /p53 fl/fl Myh6Cre +/− mice. Bar=10 µm. ( K ) LV cardiomyocyte cross‐sectional area from Ctl (n=5), Mybpc3 −/− (n=5), and Mybpc3 −/− /p53 fl/fl Myh6Cre +/− (n=5) mice. Minimum 50 cells/sample measured. All results are shown as mean±SEM.

Article Snippet: Genomic DNA was extracted from snap‐frozen P7 myocardial tissue, according to commercially available DNA extraction kit (Qiagen, 69504).

Techniques: Generated, Western Blot, Expressing, Immunohistochemical staining, Staining

Journal: iScience

Article Title: Genomic characteristics of emerging human pathogen Rickettsia aeschlimannii isolated from two Hyalomma tick species

doi: 10.1016/j.isci.2025.112080

Figure Lengend Snippet:

Article Snippet: MiniBEST Universal Genomic DNA Extraction Kit , Takara , Cat#9765.

Techniques: Virus, Recombinant, Marker, Modification, Saline, Staining, Extraction, DNA Extraction, Software, Bacteria