Journal: iScience

Article Title: Ultra-large scale virtual screening identifies a small molecule inhibitor of the Wnt transporter Wntless

doi: 10.1016/j.isci.2024.110454

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Article Snippet: 40 μL of protein A/G plus agarose (#sc-2003, Santa Cruz) was added the next day and incubated for another 2 h before washing and elution.

Techniques: Recombinant, Protease Inhibitor, Western Blot, Luciferase, Bicinchoninic Acid Protein Assay, Knock-Out, Software

Journal: eLife

Article Title: Iron derived from autophagy-mediated ferritin degradation induces cardiomyocyte death and heart failure in mice

doi: 10.7554/eLife.62174

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Article Snippet: Precleared homogenates were subjected to immunoprecipitation using 1 μg of the anti-NCOA4 antibody (Sigma-Aldrich, SAB1404569) or mouse immunoglobulin G (IgG; Santa Cruz biochemistry, sc-2025) and 40 μL of magnetic beads-coupled protein G at 4 °C for 2 hr.

Techniques: Western Blot, Immunofluorescence, Diagnostic Assay, Magnetic Beads, SYBR Green Assay, Staining, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Iron Assay, Multiple Displacement Amplification, Glutamate Assay, Electrophoretic Mobility Shift Assay, PCR Cloning, Software

Journal: Nature Communications

Article Title: Mechanically activated snai1b coordinates the initiation of myocardial delamination for trabeculation

doi: 10.1038/s41467-025-62285-w

Figure Lengend Snippet: a , b At 14 dpf, the expression of myocardial snai1b (dashed green outline) remained sparse in the ventricle, whereas Isoproterenol (ISO) treatment from 1 to 11 dpf revealed a persistent snai1b activation in the trabecular network. Anatomic labels: BA bulbus arteriosus, V ventricle, BV bulbus-ventricular annulus, Myo myocardium, Endo endocardium, Epi epicardium, Ery erythrocyte. c At 4 dpf, 4-D mapping of myocardial strain in a control and an ISO-treated heart. Two time points during diastole were displayed, and the ISO-treated heart experienced higher strain at the end-diastole time point than the control. Red dashed lines and squares mark the cross-section planes, and the red arrows indicate the viewing direction. Blue and green arrows indicate the direction of blood flow. Anatomic labels: V ventricle, AV atrioventricular canal, OFT outflow tract. d Average epicardial/endocardial ventricular strain within the sampling regions during one cardiac cycle. ISO treatment increased the myocardial strain across the endocardial surface during the end-diastolic and systolic phases. Source data are provided as a file. e Strain variations within the sampling regions, calculated as the standard deviation of strains at each time point (phase). ISO treatment induced greater myocardial strain variations compared with the control heart. Source data are provided as a file.

Article Snippet: A spinning-disk confocal microscope was used to capture 3D+time images of the ventricular myocardium (scanning unit: Yokogawa CSU-X1; microscope: Leica DMI 6000B; camera: Andor iXon plus 897; objectives: Leica HC PL IRAPO 40× or 20× water immersion lens; control software: Micro-Manager 2.0 with custom control code).

Techniques: Expressing, Activation Assay, Control, Sampling, Standard Deviation

Journal: Nature Communications

Article Title: Mechanically activated snai1b coordinates the initiation of myocardial delamination for trabeculation

doi: 10.1038/s41467-025-62285-w

Figure Lengend Snippet: a – d Abundant col1a2 mRNA colocalized with snai1b mRNA in epicardial, valvular cells (red arrowheads), and bulbus smooth muscle cells (empty red arrowheads). However, col1a2 expression was not observed in snai1b + CMs (white arrowheads) of control ( a ), ISO-treated ( b ), and CRISPRa-injected ( d ) hearts at 4 dpf. Number of hearts: Control = 10, ISO = 6, CRISPRa (VPR) = 10. e – h At 3 dpf, LY364947 , a TGF-β signaling inhibitor, attenuated trabeculation (white outlines), but the expression of snai1b (arrowheads) within BV annulus and trabecular CMs persisted. Number of hearts: Control n = 10, LY364947 n = 12. BA bulbus arteriosus, V ventricle, BV bulbus-ventricular annulus, A atrium, Ch chest wall, Myo myocardium, AV atrioventricular canal.

Article Snippet: A spinning-disk confocal microscope was used to capture 3D+time images of the ventricular myocardium (scanning unit: Yokogawa CSU-X1; microscope: Leica DMI 6000B; camera: Andor iXon plus 897; objectives: Leica HC PL IRAPO 40× or 20× water immersion lens; control software: Micro-Manager 2.0 with custom control code).

Techniques: Expressing, Control, Injection