Sapropterin reduces coronary artery malformation in offspring of pregestational diabetes mice

Maternal nicotine exposure induces congenital heart defects in the offspring of mice

“…E18.5 foetal heart sections were immunostained using anti‐α‐smooth muscle actin primary antibody (1:3000 dilution, Abcam,) to visualize coronary arteries. 31 Sections were blocked in a solution of 0.3% H 2 O 2 diluted in phosphate buffered saline (PBS) prior to incubation in secondary antibody. The primary antibody was left on overnight at room temperature in a humidity chamber.…”

“…The volume of the labelled components was calculated, and a ratio of coronary artery to myocardial volume was obtained. 31 …”

“…After 3 days, images were captured using a phase contrast microscope (Observer D1, Zeiss), and the number of spindle‐shaped cells which had grown outward from the explanted ventricles were quantified as previously described. 26 , 31 …”

“…The compact myocardium, and the right and left ventricles or coronary arteries were labelled manually in each section. The volume of the labelled components was calculated, and a ratio of coronary artery to myocardial volume was obtained 31 …”

“…M199 medium (Sigma‐Aldrich) was then added to the culture either with or without nicotine (100 ng/ml, (‐)‐nicotine N3876 Sigma‐Aldrich). After 3 days, images were captured using a phase contrast microscope (Observer D1, Zeiss), and the number of spindle‐shaped cells which had grown outward from the explanted ventricles were quantified as previously described 26,31 …”