Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Platelet Internalization Mediates Ferroptosis in Myocardial Infarction

doi: 10.1161/atvbaha.122.318161

Figure Lengend Snippet: Figure 2. Platelets are internalized by cardiomyocytes. A, Distribution of platelets in myocardial tissue (n=4). Platelets were labeled with CD41 (green), cardiomyocytes were labeled with cTnI (red), and nuclei visualized with DAPI (blue). Scale bar, 500 μm. B, Neonatal mouse cardiomyocytes (CMs) internalize platelets (n=3). CMs were labeled by CM-Dil (red), platelets were labeled by CFSE (green). Scale bar, 100 µm. C, Three-dimensional reconstruction of confocal Z-stack images of CMs co-cultured with platelets to demonstrate uptake and perinuclear localization (n=3; view angle: 30, z-Scaling: 2).

Article Snippet: Primary antibodies included cTnI antibody (1:100, Proteintech, catalog 21652-1- AP) and CD41 antibody (1:200, Santa cruz, sc-365938).

Techniques: Labeling, Cell Culture

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

Article Title: Protein Changes Contributing to Right Ventricular Cardiomyocyte Diastolic Dysfunction in Pulmonary Arterial Hypertension

doi: 10.1161/JAHA.113.000716

Figure Lengend Snippet: Sarcomeric protein phosphorylation. A, cTnI total phosphorylation was determined by ProQ‐Diamond staining, while total protein content was determined by SYPRO‐Ruby staining (n Don =10, n PAH =10). B, MyBPC total phosphorylation was determined by ProQ staining, while total protein content was determined by SYPRO staining (n Don =10, n PAH =10). C, Typical example of donor and PAH samples ProQ and SYPRO staining and the relative location of specific sarcomeric protein. ProQ staining annotations—ov, ovalbumin; β, β‐casein; MyBPC, myosin binding protein C; cTnT, cardiac troponin T; cTnI, cardiac troponin I; MLC 2 , myosin light chain 2. SYPRO staining annotations—MHC, myosin heavy chain. Data presented as mean±SEM. PAH inidcates pulmonary arterial hypertension.

Article Snippet: Blots were incubated with the following primary antibodies against specific protein or protein phosphorylation sites: cTnI dephosphorylated form (4T46, mouse monoclonal antibody, HyTest), cTnI PKA‐specific serine 22/23 site phosphorylation (4004, rabbit polyclonal antibody; Cell Signaling Technology).

Techniques: Phospho-proteomics, Staining, Binding Assay

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

Article Title: Protein Changes Contributing to Right Ventricular Cardiomyocyte Diastolic Dysfunction in Pulmonary Arterial Hypertension

doi: 10.1161/JAHA.113.000716

Figure Lengend Snippet: cTnI phosphorylation. A, Typical example of donor and PAH samples immunostained against the unphosphorylated form of cTnI and normalized for Ponceau‐S stained actin. cTnI total phosphorylation was determined by Western blot analysis in donor and PAH samples (n Don =8, n PAH =9). B, Typical example of donor and PAH samples immunostained against cTnI serine 22/23 residue and normalized for Ponceau‐S stained actin. cTnI serine 22/23 phosphorylation was determined by Western blot analysis in donor and PAH samples (n Don =9, n PAH =11). C, Typical example of donor and PAH samples immunostained against unphosphorylated (P 0 ) cTnI, mono‐phosphorylated (P 1 ) cTnI and bis‐phosphorylated (P 2 ) cTnI. cTnI relative phosphorylation distribution was determined by Phos‐Tag analysis in donor and PAH samples (n Don =5, n PAH =9). Data presented as mean±SEM. cTnI indicates cardiac troponin I; cTnT, cardiac troponin T; PAH, pulmonary arterial hypertension.

Article Snippet: Blots were incubated with the following primary antibodies against specific protein or protein phosphorylation sites: cTnI dephosphorylated form (4T46, mouse monoclonal antibody, HyTest), cTnI PKA‐specific serine 22/23 site phosphorylation (4004, rabbit polyclonal antibody; Cell Signaling Technology).

Techniques: Phospho-proteomics, Staining, Western Blot, Residue

Journal: Scientific reports

Article Title: Human platelet lysate combined with mesenchymal stem cells pretreated with platelet lysate improved cardiac function in rats with myocardial infarction.

doi: 10.1038/s41598-024-79050-6

Figure Lengend Snippet: Fig. 9. Immunofluorescent analysis indicates the expression of NKX2.5 and cTnI (green) in the hearts of rats with MI two and four weeks after PMSCs and HPL + PMSCs injection: (A, B): Sample micrographs of NKX2.5 and cTnI in an animal of each group. Quantification of the levels of NKX2.5 + and cTnI+ (C and E) and the number of Dil+/NKX2.5 + and Dil+/ cTnI+ (D and F) in the study groups. Stem cells stained with Dil dye (red) and nuclei with DAPI (blue). *** P < 0.001 vs. PMSCs 2 weeks; # < P < 0.05, ### P < 0.001 vs. HPL + PMSCs 2 weeks; &&& P < 0.01 vs. PMSCs; $$ P < 0.001, $$$ P < 0.001 vs. HPL + PMSCs. n = 3 in each group.

Article Snippet: Rabbit polyclonal primary antibody for NKX2 (No: orb540657), rabbit polyclonal antibody for cTnI (No: orb304638), and goat anti-rabbit IgG (H + L) antibody (FITC) (No orb688925) were purchased from Biorbyt, United Kingdom.

Techniques: Expressing, Injection, Staining