Cell Culture:Article Title: Metabolic alterations of endothelial cells under transient and persistent hypoxia: study using a 3D microvessels-on-chip model.
Article Snippet: Numerous signaling pathways are activated during hypoxia to facilitate angiogenesis, promoting interactions among endothelial cells and initiating downstream signaling cascades.. Although the pivotal role of the nitric oxide (NO) response pathway is well-established, the involvement of arginine-specific metabolism and bioactive lipid mechanisms in 3D flow-activated in vitro models remains less understood.. In this study, we explored the levels of arginine-specific metabolites and bioactive lipids in human coronary artery endothelial cells (HCAECs) under both transient and persistent hypoxia.
Article Title: Enabling high-sensitivity live single-cell mass spectrometry using an integrated electrical lysis and nano electrospray ionization interface.
Article Snippet: .. HCAECs (PromoCell, C-12221) were resuspended in 10 ml fresh EGM MV2 medium with supplements (PromoCell, C-22022, C39216) and cultured in T75 flasks (Nunc Easy flask, Sigma, F7552). ..
Article Title: CXCL10-dependent epithelial-vascular cross-talk for endothelial activation following SARS-CoV-2 infection
Article Snippet: Vero-E6 (ATCC, CRL-1586) and Calu-3 cells (ATCC, HTB-55) were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Gibco) supplemented with 10% fetal calf serum and gentamicin (50 μg/mL) at 37 °C in a humidified 5% CO 2 incubator. .. HUVECs (Promocell, C-12203), HUAECs (Promocell, C-12202), and HCAECs (Promocell, C-12221) were maintained in endothelial cell growth medium 2 (EGM2; C-22111, PromoCell) in a cell culture dish coated with 0.5% gelatin solution. .. Human monocytic THP-1 cells (ATCC, TIB-202) were cultured in RPMI-1640 medium (Life Technologies) supplemented with 10% FBS, 2 mM L-glutamine and 1% penicillin and streptomycin.
Article Title: Metabolic alterations of endothelial cells under transient and persistent hypoxia: study using a 3D microvessels-on-chip model
Article Snippet: .. HCAECs (PromoCell, C-12221) were resuspended in 10 ml fresh EGM MV2 medium with supplements (PromoCell, C-22022, C-39216) and cultured in T75 flasks (Nunc Easyflask, Sigma, F7552). ..
Article Title: CXCL10-dependent epithelial-vascular cross-talk for endothelial activation following SARS-CoV-2 infection.
Article Snippet: Vero-E6 (ATCC, CRL-1586) and Calu-3 cells (ATCC, HTB-55) were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Gibco) supplemented with 10% fetal calf serum and gentamicin (50 μg/mL) at 37 °C in a Scientific Reports | (2025) 15:21129 10| https://doi.org/10.1038/s41598-025-08329-z humidified 5% CO2 incubator. .. HUVECs (Promocell, C-12203), HUAECs (Promocell, C-12202), and HCAECs (Promocell, C-12221) were maintained in endothelial cell growth medium 2 (EGM2; C-22111, PromoCell) in a cell culture dish coated with 0.5% gelatin solution. .. Human monocytic THP-1 cells (ATCC, TIB-202) were cultured in RPMI-1640 medium (Life Technologies) supplemented with 10% FBS, 2 mM L-glutamine and 1% penicillin and streptomycin.
Incubation:Article Title: RV-specific Targeting of Snai1 Rescues Pulmonary Hypertension-induced Right Ventricular Failure by Inhibiting EndMT and Fibrosis via LOXL2 Mediated Mechanism
Article Snippet: All images were acquired using a confocal microscope (Nikon). .. HCAECs: Human coronary artery endothelial cells (HCAECs: Promocell# C-12221) were incubated in hypoxia condition in the hypoxia chamber with TGFβ1 (recombinant human TGFβ1: 15ng/ml, Cat #T7039, Sigma) for 72 h. Snai1-KD was performed in HCAECs under hypoxia and TGFβ1 (15ng/ml) treatment using Human SNAI1 siRNA [1uM/ml of medium of in vitro ready Accell Human SNAI1siRNA-SMARTpool (6615), and appropriate scramble control (siScrm; D-001910-01-50) commercially available from Horizon Discovery, Lafayette, CO, USA] in combination with the LipofectamineTM RNAiMAX Transfection Reagent (Cat# 13778100, Invitrogen) for enhanced transfection efficiency, following the manufacturer’s instructions. qPCR: Total RNA was isolated from the HCAECs using RNeasy Mini Kit (Cat #74104 and 74106, QIAGEN). .. Quantitative real-time PCR was performed following the protocol for the “mixed primer strategy” (Random primer + Oligo dT) using the SsoAdvanced Universal SYBR Green Supermix (BioRad, Cat #172-5274) in the CFX ConnectTM Real-Time PCR Detection System (384-well block, Biorad, Cat #1855201).
Recombinant:Article Title: RV-specific Targeting of Snai1 Rescues Pulmonary Hypertension-induced Right Ventricular Failure by Inhibiting EndMT and Fibrosis via LOXL2 Mediated Mechanism
Article Snippet: All images were acquired using a confocal microscope (Nikon). .. HCAECs: Human coronary artery endothelial cells (HCAECs: Promocell# C-12221) were incubated in hypoxia condition in the hypoxia chamber with TGFβ1 (recombinant human TGFβ1: 15ng/ml, Cat #T7039, Sigma) for 72 h. Snai1-KD was performed in HCAECs under hypoxia and TGFβ1 (15ng/ml) treatment using Human SNAI1 siRNA [1uM/ml of medium of in vitro ready Accell Human SNAI1siRNA-SMARTpool (6615), and appropriate scramble control (siScrm; D-001910-01-50) commercially available from Horizon Discovery, Lafayette, CO, USA] in combination with the LipofectamineTM RNAiMAX Transfection Reagent (Cat# 13778100, Invitrogen) for enhanced transfection efficiency, following the manufacturer’s instructions. qPCR: Total RNA was isolated from the HCAECs using RNeasy Mini Kit (Cat #74104 and 74106, QIAGEN). .. Quantitative real-time PCR was performed following the protocol for the “mixed primer strategy” (Random primer + Oligo dT) using the SsoAdvanced Universal SYBR Green Supermix (BioRad, Cat #172-5274) in the CFX ConnectTM Real-Time PCR Detection System (384-well block, Biorad, Cat #1855201).
In Vitro:Article Title: RV-specific Targeting of Snai1 Rescues Pulmonary Hypertension-induced Right Ventricular Failure by Inhibiting EndMT and Fibrosis via LOXL2 Mediated Mechanism
Article Snippet: All images were acquired using a confocal microscope (Nikon). .. HCAECs: Human coronary artery endothelial cells (HCAECs: Promocell# C-12221) were incubated in hypoxia condition in the hypoxia chamber with TGFβ1 (recombinant human TGFβ1: 15ng/ml, Cat #T7039, Sigma) for 72 h. Snai1-KD was performed in HCAECs under hypoxia and TGFβ1 (15ng/ml) treatment using Human SNAI1 siRNA [1uM/ml of medium of in vitro ready Accell Human SNAI1siRNA-SMARTpool (6615), and appropriate scramble control (siScrm; D-001910-01-50) commercially available from Horizon Discovery, Lafayette, CO, USA] in combination with the LipofectamineTM RNAiMAX Transfection Reagent (Cat# 13778100, Invitrogen) for enhanced transfection efficiency, following the manufacturer’s instructions. qPCR: Total RNA was isolated from the HCAECs using RNeasy Mini Kit (Cat #74104 and 74106, QIAGEN). .. Quantitative real-time PCR was performed following the protocol for the “mixed primer strategy” (Random primer + Oligo dT) using the SsoAdvanced Universal SYBR Green Supermix (BioRad, Cat #172-5274) in the CFX ConnectTM Real-Time PCR Detection System (384-well block, Biorad, Cat #1855201).
Control:Article Title: RV-specific Targeting of Snai1 Rescues Pulmonary Hypertension-induced Right Ventricular Failure by Inhibiting EndMT and Fibrosis via LOXL2 Mediated Mechanism
Article Snippet: All images were acquired using a confocal microscope (Nikon). .. HCAECs: Human coronary artery endothelial cells (HCAECs: Promocell# C-12221) were incubated in hypoxia condition in the hypoxia chamber with TGFβ1 (recombinant human TGFβ1: 15ng/ml, Cat #T7039, Sigma) for 72 h. Snai1-KD was performed in HCAECs under hypoxia and TGFβ1 (15ng/ml) treatment using Human SNAI1 siRNA [1uM/ml of medium of in vitro ready Accell Human SNAI1siRNA-SMARTpool (6615), and appropriate scramble control (siScrm; D-001910-01-50) commercially available from Horizon Discovery, Lafayette, CO, USA] in combination with the LipofectamineTM RNAiMAX Transfection Reagent (Cat# 13778100, Invitrogen) for enhanced transfection efficiency, following the manufacturer’s instructions. qPCR: Total RNA was isolated from the HCAECs using RNeasy Mini Kit (Cat #74104 and 74106, QIAGEN). .. Quantitative real-time PCR was performed following the protocol for the “mixed primer strategy” (Random primer + Oligo dT) using the SsoAdvanced Universal SYBR Green Supermix (BioRad, Cat #172-5274) in the CFX ConnectTM Real-Time PCR Detection System (384-well block, Biorad, Cat #1855201).
Transfection:Article Title: RV-specific Targeting of Snai1 Rescues Pulmonary Hypertension-induced Right Ventricular Failure by Inhibiting EndMT and Fibrosis via LOXL2 Mediated Mechanism
Article Snippet: All images were acquired using a confocal microscope (Nikon). .. HCAECs: Human coronary artery endothelial cells (HCAECs: Promocell# C-12221) were incubated in hypoxia condition in the hypoxia chamber with TGFβ1 (recombinant human TGFβ1: 15ng/ml, Cat #T7039, Sigma) for 72 h. Snai1-KD was performed in HCAECs under hypoxia and TGFβ1 (15ng/ml) treatment using Human SNAI1 siRNA [1uM/ml of medium of in vitro ready Accell Human SNAI1siRNA-SMARTpool (6615), and appropriate scramble control (siScrm; D-001910-01-50) commercially available from Horizon Discovery, Lafayette, CO, USA] in combination with the LipofectamineTM RNAiMAX Transfection Reagent (Cat# 13778100, Invitrogen) for enhanced transfection efficiency, following the manufacturer’s instructions. qPCR: Total RNA was isolated from the HCAECs using RNeasy Mini Kit (Cat #74104 and 74106, QIAGEN). .. Quantitative real-time PCR was performed following the protocol for the “mixed primer strategy” (Random primer + Oligo dT) using the SsoAdvanced Universal SYBR Green Supermix (BioRad, Cat #172-5274) in the CFX ConnectTM Real-Time PCR Detection System (384-well block, Biorad, Cat #1855201).
Real-time Polymerase Chain Reaction:Article Title: RV-specific Targeting of Snai1 Rescues Pulmonary Hypertension-induced Right Ventricular Failure by Inhibiting EndMT and Fibrosis via LOXL2 Mediated Mechanism
Article Snippet: All images were acquired using a confocal microscope (Nikon). .. HCAECs: Human coronary artery endothelial cells (HCAECs: Promocell# C-12221) were incubated in hypoxia condition in the hypoxia chamber with TGFβ1 (recombinant human TGFβ1: 15ng/ml, Cat #T7039, Sigma) for 72 h. Snai1-KD was performed in HCAECs under hypoxia and TGFβ1 (15ng/ml) treatment using Human SNAI1 siRNA [1uM/ml of medium of in vitro ready Accell Human SNAI1siRNA-SMARTpool (6615), and appropriate scramble control (siScrm; D-001910-01-50) commercially available from Horizon Discovery, Lafayette, CO, USA] in combination with the LipofectamineTM RNAiMAX Transfection Reagent (Cat# 13778100, Invitrogen) for enhanced transfection efficiency, following the manufacturer’s instructions. qPCR: Total RNA was isolated from the HCAECs using RNeasy Mini Kit (Cat #74104 and 74106, QIAGEN). .. Quantitative real-time PCR was performed following the protocol for the “mixed primer strategy” (Random primer + Oligo dT) using the SsoAdvanced Universal SYBR Green Supermix (BioRad, Cat #172-5274) in the CFX ConnectTM Real-Time PCR Detection System (384-well block, Biorad, Cat #1855201).
Isolation:Article Title: RV-specific Targeting of Snai1 Rescues Pulmonary Hypertension-induced Right Ventricular Failure by Inhibiting EndMT and Fibrosis via LOXL2 Mediated Mechanism
Article Snippet: All images were acquired using a confocal microscope (Nikon). .. HCAECs: Human coronary artery endothelial cells (HCAECs: Promocell# C-12221) were incubated in hypoxia condition in the hypoxia chamber with TGFβ1 (recombinant human TGFβ1: 15ng/ml, Cat #T7039, Sigma) for 72 h. Snai1-KD was performed in HCAECs under hypoxia and TGFβ1 (15ng/ml) treatment using Human SNAI1 siRNA [1uM/ml of medium of in vitro ready Accell Human SNAI1siRNA-SMARTpool (6615), and appropriate scramble control (siScrm; D-001910-01-50) commercially available from Horizon Discovery, Lafayette, CO, USA] in combination with the LipofectamineTM RNAiMAX Transfection Reagent (Cat# 13778100, Invitrogen) for enhanced transfection efficiency, following the manufacturer’s instructions. qPCR: Total RNA was isolated from the HCAECs using RNeasy Mini Kit (Cat #74104 and 74106, QIAGEN). .. Quantitative real-time PCR was performed following the protocol for the “mixed primer strategy” (Random primer + Oligo dT) using the SsoAdvanced Universal SYBR Green Supermix (BioRad, Cat #172-5274) in the CFX ConnectTM Real-Time PCR Detection System (384-well block, Biorad, Cat #1855201).
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