Journal: Translational Research

Article Title: Fibrotic extracellular matrix impacts cardiomyocyte phenotype and function in an iPSC-derived isogenic model of cardiac fibrosis

doi: 10.1016/j.trsl.2024.07.003

Figure Lengend Snippet: List of primary antibodies.

Article Snippet: , Cardiac TNNT-FITC , 130-119-575 , 1:50 , Miltenyi Biotec.

Techniques: Immunofluorescence, Flow Cytometry

Journal: Advanced Science

Article Title: PICALM Regulating the Generation of Amyloid β‐Peptide to Promote Anthracycline‐Induced Cardiotoxicity

doi: 10.1002/advs.202401945

Figure Lengend Snippet: Amyloid β metabolic process pathway and related genes were the major issue in DOX‐induced cardiotoxicity mice and patients with chemotherapy‐induced cardiotoxicity. A) Bar plot for scGSVA analysis of CMs. B) Heat plot of the associations between genes and pathways. C) Violin plots showing the expression of Lpin1 / Psap / Gnptab / App in each cardiomyocyte cluster. D,E) Representative and statistical graphs of APP expression at the myocardial cells in DOX‐induced cardiotoxicity group and patients. Scan bars, 50 µm; scale bars for the magnified images, 20 µm. F) Diagram showing the potential mechanisms of DOX induction in CMs. G,H) Representative and statistical graphs of Aβ40, PICALM expression at the CMs marked by TNNT2 in DOX‐induced cardiotoxicity mice, and a diagram showing the correlation of PICALM and Aβ40 in mice CMs. Scan bars, 1000 µm; scale bars for the magnified images, 200 µm. I,J) Representative and statistical graphs of Aβ40, PICALM expression at the CMs marked by TNNT2 in patients with chemotherapy‐induced cardiotoxicity, and a diagram showing the correlation of PICALM and Aβ40 in human CMs. Scan bars, 500 µm; scale bars for the magnified images, 50 µm. K) ELISA analysis of mouse serum Aβ40 level in DOX‐induced cardiotoxicity. L) ELISA analysis of human serum Aβ40 level in chemotherapy‐induced cardiotoxicity and plasma Aβ40 levels could distinguish CICP from non‐CICP (AUC = 0.94, P = 0.03). NC, normal control; DOX, DOX‐treated mice; CICP, chemotherapy‐induced cardiotoxicity patients. Mice: n = 5/group in RNA‐seq and n = 8/group in immunoimaging; human: n = 3/group. Student's t‐test was applied to analyze the differences between two groups. Multiple‐group comparisons were made by one‐way ANOVA followed by the Tukey test. * P < 0.05; ** P < 0.01; *** P < 0.001. Data are presented as the mean ± SD.

Article Snippet: After antigen repair, closure, and permeability, the primary antibody (anti‐PICALM; anti‐TNNT; anti‐LPIN1, bs‐0759R, bioss; anti‐APP, bs‐0112 M, bioss; anti‐Aβ40) incubation, sections were incubated with a secondary antibody with Alexa Fluor Dye (Thermo Fisher Scientific).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Control, RNA Sequencing Assay

Journal: Advanced Science

Article Title: PICALM Regulating the Generation of Amyloid β‐Peptide to Promote Anthracycline‐Induced Cardiotoxicity

doi: 10.1002/advs.202401945

Figure Lengend Snippet: Si‐Picalm can alleviate DOX‐induced hiPSC‐CMs damage and Aβ peptide production. A) Cell viability in different groups. Cells were treated with the indicated concentration of DOX (0–10 µm). After incubation, cell viability was measured by CCK8 assay. B,C) Statistical analysis of vesicle area, PICALM and Aβ40 level of hiPSC‐CMs treated with 1 µM DOX for 0, 12, 24, and 36 h with confocal microscope (Zeiss) by using ImageJ plugins. [ ²³ ] D) The statistical graphs determined using an IncuCyte imaging system. H9c2 cells pre‐stained with 0.5 µM LysoTracker Red for 30 min were treated with 1 µm DOX for 24 h. E) The statistical graphs of LPIN1 expression determined by confocal. F) The WGA staining of DOX‐treated cells and statistical graphs of vesicles area. G) Representative images of ROS and calcium concentration of hiPSC‐CMs treated with DOX or Aβ40 and statistical analysis. Scan bars, 50 µm. H) Representative images of propidium iodide (PI)‐staining of hiPSC‐CMs treated with DOX or Aβ40 and statistical analysis of apoptosis and necrotic cell percentage. Scan bars, 200 µm. I) Statistical analysis of Aβ40 level of hiPSC‐CMs using ELISA in the medium. J) The western blot of the hiPSC‐CMs groups including SI‐NC, SI‐PICALM, DOX+SI‐NC, DOX+SI‐PICALM, and statistical graphs of PICALM, APP, and Aβ40 expression. K,L) Representative images and statistical analysis of WGA/LPIN1 staining in the hiPSC‐CMs groups. Scan bars, 200 µm. M) Representative images and statistical analysis of lysosome tracking in the hiPSC‐CMs groups. Scan bars, 50 µm. N) The cell viability in groups. O,P) Representative images and statistical analysis of TUNEL assay in the hiPSC‐CMs groups. Scan bars, 200 µm. DOX, DOX‐treated cells; SI‐NC, SI‐GFP; OE‐NC, overexpression vector. Student's t‐test was applied to analyze the differences between two groups. Multiple‐group comparisons were made by one‐way ANOVA followed by the Tukey test. * P < 0.05; ** P < 0.01; *** P < 0.001. Data are presented as the mean ± SD.

Article Snippet: After antigen repair, closure, and permeability, the primary antibody (anti‐PICALM; anti‐TNNT; anti‐LPIN1, bs‐0759R, bioss; anti‐APP, bs‐0112 M, bioss; anti‐Aβ40) incubation, sections were incubated with a secondary antibody with Alexa Fluor Dye (Thermo Fisher Scientific).

Techniques: Concentration Assay, Incubation, CCK-8 Assay, Microscopy, Imaging, Staining, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, TUNEL Assay, Over Expression, Plasmid Preparation