C 12285

Primary human cardiac microvascular endothelial cells (HCMECs) (C-12285), EC growth medium MV, detachKit, and cryo-SFM were obtained from PromoCell, Inc. (Germany). Cells were cultured in EC growth medium MV without antibiotics or antimycotics and gently detached using a detachKit. HCMECs at passages 5–7 were maintained in an incubator (37 °C, 5 % CO₂) until they reached a cell density of approximately 80 % confluency, prior to subsequent experiments.

Cardiac microvascular endothelial cells (CMECs; C-12285, PromoCell) were cultured in endothelial cell growth medium MV (C-22020, PromeCell) supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin and 100 µg/ml streptomycin at 37°C in a 5% CO₂ humidified incubator. Ang II (100 µM, 24 h) was used to induce the in vitro HP cell model as previously described (17 (link),18 (link)). Mst1 silencing was achieved by transfecting Ad-sh-Mst1 [MOI: 200; Hanheng Biotechnology (Shanghai) Co., Ltd.] into the cells. The cultured CMECs were treated with 50 nM bafilomycin A1 (MiilliporeSigma) for 2 h to evaluate the autophagic flux.