Journal: Cells

Article Title: HDAC6 and ERK/ADAM17 Regulate VEGF-Induced NOTCH Signaling in Lung Endothelial Cells

doi: 10.3390/cells12182231

Figure Lengend Snippet: VEGF induces rapid NICD expression through ERK1/2. ( A ) NICD expression was assessed 30 min after VEGF treatment in HPMEC-Im, as shown by immunoblotting, with densitometry shown graphically; ( B ) n = 3 independent experiments, * p < 0.05, VEGF vs. Ctrl; ( C ) mRNA expression of DLL4 , HES1 , HEY1/2 , and NRARP was quantified by qRT-PCR after transfection of empty or NICD-contained plasmid in HPMEC-Im, n = 10 independent experiments; * p < 0.05, empty plasmid vs. NICD overexpression (OE). ( D ) HPMEC-Im were treated with 50 μg/mL VEGF for 30 min with or without U0126 (ERK inhibitor) pretreatment, and DLL4, NICD, ERK, and p-ERK expression were quantified by immunoblotting, with densitometry analysis presented graphically ( E ), n = 5 independent experiments, * p < 0.05, VEGF vs. Ctrl. ( F ) DLL4 and HES1 mRNA expression was assessed by qRT-PCR 5 h after 50 μg/mL VEGF treatment. n = 9 independent experiments, * p < 0.05.

Article Snippet: Cell culture and reagents: Embryonic human pulmonary microvascular endothelial cells (HPMEC) were purchased from ScienCell (Carlsbad, CA, USA) and were immortalized (HPMEC-Im) with SV40 large T antigen transformation as described before by our lab [ ].

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Transfection, Plasmid Preparation, Over Expression

Journal: Cells

Article Title: HDAC6 and ERK/ADAM17 Regulate VEGF-Induced NOTCH Signaling in Lung Endothelial Cells

doi: 10.3390/cells12182231

Figure Lengend Snippet: VEGF induces rapid NICD expression through DLL4, ADAM17, and ɣ-secretase. NICD expression was quantified by immunoblotting 30 min after VEGF treatment in HPMEC-Im ( A ), with densitometry analysis shown graphically ( B ), n = 4 independent experiments, * p < 0.05, ns means no significance. HPMEC-Im transduced with lentivirus containing sc shRNA or DLL4 shRNA were treated with VEGF, and DLL4 , HES1 , and HEY1 mRNA expression was assessed by qRT-PCR ( C ), n = 6 independent experiments, * p < 0.05. NICD protein expression was assessed by immunoblotting after VEGF treatment with or without KP457 pretreatment ( D ). Densitometry analysis is presented graphically ( E ), n = 5 independent experiments, * p < 0.05. HPMEC-Im were treated with VEGF with or without KP457 pretreatment, and HES1 and HEY1 mRNA expression was assessed by qRT-PCR ( F ), n = 9 independent experiments, * p < 0.05. NICD protein expression was assessed by immunoblotting after VEGF treatment with or without DBZ pretreatment ( G ). Densitometry analysis is presented graphically ( H ), n = 3 independent experiments, * p < 0.05.

Article Snippet: Cell culture and reagents: Embryonic human pulmonary microvascular endothelial cells (HPMEC) were purchased from ScienCell (Carlsbad, CA, USA) and were immortalized (HPMEC-Im) with SV40 large T antigen transformation as described before by our lab [ ].

Techniques: Expressing, Western Blot, Transduction, shRNA, Quantitative RT-PCR

Journal: Cells

Article Title: HDAC6 and ERK/ADAM17 Regulate VEGF-Induced NOTCH Signaling in Lung Endothelial Cells

doi: 10.3390/cells12182231

Figure Lengend Snippet: Inhibition of HDAC6 leads to increased expression of NICD but suppresses the Notch signaling pathway. HPMEC-Im transduced with lentivirus containing sc shRNA or HDAC6 shRNA. ( A ) DLL4 , HES1 , HEY1, and JAG1 mRNA expression was assessed by qRT-PCR sc shRNA vs. HDAC6 shRNA, n = 6 independent experiments, * p < 0.05. ( B ) NICD protein expression was assessed by immunoblotting, with densitometry shown graphically ( C ), n = 4 independent experiments, and * p < 0.05. ( D ) Acetylated tubulin expression and ERK phosphorylation were assessed by immunoblotting, with densitometry shown graphically ( E ), n = 3 independent experiments, and * p < 0.05. ( F ) TP1-Luc2 luciferase assay was applied to assess Notch signaling activity in sc shRNA or HDAC6 shRNA-expressed HEK293 cells, n = 5 independent experiments and * p < 0.05.

Article Snippet: Cell culture and reagents: Embryonic human pulmonary microvascular endothelial cells (HPMEC) were purchased from ScienCell (Carlsbad, CA, USA) and were immortalized (HPMEC-Im) with SV40 large T antigen transformation as described before by our lab [ ].

Techniques: Inhibition, Expressing, Transduction, shRNA, Quantitative RT-PCR, Western Blot, Phospho-proteomics, Luciferase, Activity Assay

Journal: Cells

Article Title: HDAC6 and ERK/ADAM17 Regulate VEGF-Induced NOTCH Signaling in Lung Endothelial Cells

doi: 10.3390/cells12182231

Figure Lengend Snippet: Regulation of NICD and SNW1 binding via HDAC6-mediated deacetylation. HPMEC-Im transduced with lentivirus containing sc shRNA or HDAC6 shRNA were treated with 50 ng/mL VEGF for 45 min. Immunoprecipitation (IP) with NICD antibody was performed with subsequent Western blot analysis carried out with an acetylated antibody and HDAC6 antibody on immunoprecipitated protein to quantify changes in NICD acetylation and HDAC6-NICD binding ( A ). Densitometric quantification of Western blot images in A is shown graphically ( B ), n = 5 independent experiments; * p < 0.05. Similarly, NICD acetylation and NICD-SNW1 binding were assessed after IP with NICD antibody ( C ), with the results presented graphically ( D ), n ≥ 4 independent experiments, * p < 0.05. SNW1 acetylation and NICD-SNW1 binding were measured through immunoblotting after IP with SNW1 antibody ( E ), and densitometry analysis shown graphically ( F ), n = 5 independent experiments and * p < 0.05. ( G ) SNW1 binding to RBPJ binding sites on the promoters of HES1 and HEY2 was quantified using Chromatin IP (ChIP) with SNW1 antibody. n = 4 independent experiments, * p < 0.05.

Article Snippet: Cell culture and reagents: Embryonic human pulmonary microvascular endothelial cells (HPMEC) were purchased from ScienCell (Carlsbad, CA, USA) and were immortalized (HPMEC-Im) with SV40 large T antigen transformation as described before by our lab [ ].

Techniques: Binding Assay, Transduction, shRNA, Immunoprecipitation, Western Blot, Chromatin Immunoprecipitation

Journal: Cells

Article Title: HDAC6 and ERK/ADAM17 Regulate VEGF-Induced NOTCH Signaling in Lung Endothelial Cells

doi: 10.3390/cells12182231

Figure Lengend Snippet: HDAC6 regulates pulmonary EC angiogenesis ( A – D ). - 3D angiogenesis was performed using a validated bead assay as described in the methods section in HPMEC-Im and treated with sc shRNA and HDAC6 shRNA ( A ). Magnified image of a representative sample from HPMEC-Im treated with sc shRNA showing an angiogenic tube with lumen ( B ). Quantification of roots, branches ( C ), and length of roots ( D ) is shown; 1-3 representative beads/well from 8 (sc shRNA) and 13 (HDAC6 shRNA) wells were evaluated for 3D angiogenesis. Data is derived from two independent experiments. (* p < 0.05). ( E – I ) Mouse lungs were inflation-fixed on P14 after three doses of HDAC6-I ip injection. CD31 immunohistochemical staining was performed ( E ) (scale bar: 50 µm) and H&E staining was performed ( G ) (scale bar: 200 µm) with mouse lung sections. Arrows in figure E show single vs. double layer capillaries in and around alveolar septum. Radial alveolar counts (RAC) ( G ) and mean linear intercepts (MLI) ( H ) graphically presented. The results were obtained from at least four mice per group, indicating statistical significance (* p < 0.05). ( I ) Western blot depicting acetylated tubulin from clarified mouse whole lung lysates.

Article Snippet: Cell culture and reagents: Embryonic human pulmonary microvascular endothelial cells (HPMEC) were purchased from ScienCell (Carlsbad, CA, USA) and were immortalized (HPMEC-Im) with SV40 large T antigen transformation as described before by our lab [ ].

Techniques: shRNA, Derivative Assay, Injection, Immunohistochemical staining, Staining, Western Blot

Journal: The American Journal of Pathology

Article Title: Pulmonary Endothelial Protein Kinase C-Delta (PKCδ) Regulates Neutrophil Migration in Acute Lung Inflammation

doi: 10.1016/j.ajpath.2013.09.010

Figure Lengend Snippet: Role of PKCδ in neutrophil transmigration. A: Selective depletion of PKCδ by stealth PKCδ siRNA in PMVECs. Controls were PMVECs treated with siRNA with equivalent percentage of GC nucleotide content (GC control) as the stealth PKCδ siRNA. Levels of specific PKC isotypes were determined in cell lysates by immunoblotting with isotype-specific antibodies to PKCδ, PKCα, PKCβI/II, and PKCζ. Blots are representative of three separate Western blotting experiments. B: Neutrophil transmigration through IL-1β–activated PMVECs. PMVECs were grown on Transwell inserts and incubated with IL-1β or buffer after transfection with PKCδ stealth siRNA or siRNA containing equivalent percentage of GC nucleotide content (Control). In a second series of experiments, PMVECs were grown on Transwell inserts and were pretreated with buffer, 10 U/mL IL-1β+vehicle, IL-1β+TAT-TAT control peptide, or IL-1β+PKCδ inhibitor. Neutrophils (1 × 106 per well) were added to the upper well of the Transwell culture system and allowed to migrate for 1.5 hours through PMVECs. C: Role of PKCδ in neutrophil transmigration mediated by IL-8 and fMLP. Chemoattractants fMLP (1 nmol/L) or IL-8 (2 nmol/L) were added to the bottom well of a Transwell culture system. Neutrophils (2 × 106 per well) were added to the upper well and were allowed to migrate for 1.5 hours through PMVECs pretreated with buffer or with 2 μmol/L PKCδ inhibitor. Data are expressed as means ± SEM. n = 4 (B, siRNA experiments); n = 3–9 (B, PKCδ inhibitor experiments); n = 6–9 (C). ∗∗P < 0.01, ∗∗∗P < 0.001 versus respective control or controls (B). ∗∗P < 0.01 fMLP versus fMLP+PKCδ inhibitor–treated PMVECs; ∗∗∗P < 0.001 fMLP or IL-8 versus buffer (C).

Article Snippet: Endothelial Cell Culture Human pulmonary microvascular endothelial cells (PMVECs) were obtained from Lonza (Walkersville, MD) and were cultured using endothelial growth medium 2 (Lonza) with added bovine brain extract, vascular endothelial growth factor, epidermal growth factor, gentamicin, and hydrocortisone according to the manufacturer’s specifications.

Techniques: Transmigration Assay, Control, Western Blot, Incubation, Transfection

Journal: The American Journal of Pathology

Article Title: Pulmonary Endothelial Protein Kinase C-Delta (PKCδ) Regulates Neutrophil Migration in Acute Lung Inflammation

doi: 10.1016/j.ajpath.2013.09.010

Figure Lengend Snippet: Neutrophil binding to IL-1β–activated PMVECs is PKCδ dependent. PMVEC monolayers were treated overnight with buffer or 10 U/mL IL-1β ± 0.5 to 10 μmol/L PKCδ inhibitor. Calcein-loaded neutrophils (2 × 105) were incubated with PMVECs for 30 minutes at 37°C. Neutrophil binding was calculated from a standard curve prepared from calcein-loaded neutrophils. Data are expressed as means ± SEM from one representative experiment, performed in triplicate, from four different neutrophil donors. ∗∗P < 0.01 IL-1β versus IL-1β+PKCδ inhibitor (1 to 10 μmol/L); ∗∗∗P < 0.001 buffer versus IL-1β.

Article Snippet: Endothelial Cell Culture Human pulmonary microvascular endothelial cells (PMVECs) were obtained from Lonza (Walkersville, MD) and were cultured using endothelial growth medium 2 (Lonza) with added bovine brain extract, vascular endothelial growth factor, epidermal growth factor, gentamicin, and hydrocortisone according to the manufacturer’s specifications.

Techniques: Binding Assay, Incubation

Journal: The American Journal of Pathology

Article Title: Pulmonary Endothelial Protein Kinase C-Delta (PKCδ) Regulates Neutrophil Migration in Acute Lung Inflammation

doi: 10.1016/j.ajpath.2013.09.010

Figure Lengend Snippet: Role of PKCδ in PMVEC adhesion molecule expression. The expression of VCAM-1, ICAM-1, and PECAM-1 on PMVEC monolayers was determined by a cell-surface ELISA. Human PMVEC monolayers were treated overnight with buffer or IL-1β ± PKCδ inhibitor (0.01 to 5 mol/L). A: IL-1β–stimulated VCAM-1 expression. Constitutive expression of VCAM-1 (buffer) was normalized to 1, and expression in response to IL-1β treatment ± PKCδ inhibitor was compared with constitutive VCAM-1 expression. B: IL-1β–stimulated ICAM-1 expression, relative to constitutive ICAM-1 expression (as described for A). C: IL-1β–stimulated PECAM-1 expression, relative to constitutive PECAM-1 expression (as described for A). Data are expressed as means ± SEM. n = 7 (A); n = 5 (B); n = 6 (C). ∗∗P < 0.01, buffer versus IL-1β–treated PMVECs, †P < 0.01, IL-1β–treated PMVECs versus 10 U/mL IL-1β+PKCδ inhibitor (0.1–5 μmol/L) (A). ∗P < 0.05, IL-1β–treated PMVECs versus IL-1β+PKCδ inhibitor (0.1 μmol/L); ∗∗P < 0.01, IL-1β–treated PMVECs versus IL-1β+PKCδ inhibitor (1–5 μmol/L); and ∗∗∗P < 0.001, buffer versus IL-1β–treated PMVECs (B).

Article Snippet: Endothelial Cell Culture Human pulmonary microvascular endothelial cells (PMVECs) were obtained from Lonza (Walkersville, MD) and were cultured using endothelial growth medium 2 (Lonza) with added bovine brain extract, vascular endothelial growth factor, epidermal growth factor, gentamicin, and hydrocortisone according to the manufacturer’s specifications.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: The American Journal of Pathology

Article Title: Pulmonary Endothelial Protein Kinase C-Delta (PKCδ) Regulates Neutrophil Migration in Acute Lung Inflammation

doi: 10.1016/j.ajpath.2013.09.010

Figure Lengend Snippet: Role for PKCδ in IL-1β–mediated ROS production and activation of the redox-sensitive transcription factor NF-κB. A: IL-1β–mediated p65 NF-κB translocation is PKCδ dependent. PMVECs were treated with buffer or 10 U/mL IL-1β ± PKCδ inhibitor. Nuclear extracts, prepared after 15 minutes of incubation, were probed for the presence of p65 NF-κB by Western blotting. Representative blots are shown, with quantitation. HDAC-2 is a marker for nuclear fractions. B: Regulatory role for ROS in IL-1β–mediated VCAM-1 expression in PMVECs. VCAM-1 expression was measured by cell-surface ELISA. Constitutive expression of VCAM-1 (buffer) was normalized to 1, and expression in response to IL-1β treatment ± the ROS inhibitors DPI (10 μmol/L) or apocynin (APO; 500 μmol/L) or appropriate vehicle controls (DMSO and EtOH, respectively) was compared. C: PKCδ inhibition attenuates ROS production in IL-1β–treated PMVECs. CM-H2DCFDA–loaded PMVEC monolayers were treated with 10 U/mL IL-1β ± 2 μmol/L PKCδ inhibitor. Controls (Dulbecco’s modified Eagle’s medium) exhibited no change in oxidation of the probe, and H2O2 confirmed presence of the probe (data not shown). Graph in C is representative of four separate experiments run in triplicate (P < 0.03 IL-1β versus IL-1β+PKCδ inhibitor). Data are expressed as means ± SEM (A and B). n = 5 (A); n = 4 (B). ∗∗∗P < 0.001 IL-1β–treated PMVECs versus buffer-treated PMVECs; †P < 0.001 IL-1β–treated PMVECs versus IL-1β+PKCδ inhibitor–treated PMVECs (A). ∗∗∗P < 0.001 buffer versus IL-1β, IL-1β+DMSO, and IL-1β+EtOH; †P < 0.001 IL-1β+DPI versus IL-1β and IL-1β+DMSO; and ∗∗P < 0.001 IL-1β+APO versus IL-1β and IL-1β+EtOH (B).

Article Snippet: Endothelial Cell Culture Human pulmonary microvascular endothelial cells (PMVECs) were obtained from Lonza (Walkersville, MD) and were cultured using endothelial growth medium 2 (Lonza) with added bovine brain extract, vascular endothelial growth factor, epidermal growth factor, gentamicin, and hydrocortisone according to the manufacturer’s specifications.

Techniques: Activation Assay, Translocation Assay, Incubation, Western Blot, Quantitation Assay, Marker, Expressing, Enzyme-linked Immunosorbent Assay, Inhibition, Modification

Journal: International Journal of Molecular Sciences

Article Title: Activation of Endothelial Large Conductance Potassium Channels Protects against TNF-α-Induced Inflammation

doi: 10.3390/ijms24044087

Figure Lengend Snippet: Effects of Ca V channel inhibition with Nifedipine and the selective BK channel opener NS1619 on inflammatory cytokine secretion: ( A , B ). The inhibition of L-type Ca V channels with Nifedipine (10 µM; 24 h) decreases the baseline and TNF-α-induced (5 ng/mL; 24 h) CCL-2 and IL-6 secretion from HULEC5a cells ( n = 5–20, * p ≤ 0.05), shown by ELISA. ( C , D ) Fluorescence intensity (IF; A.U. = arbitrary units) measured by fluorometric FLIPR assays depicts Em changes in HULEC5a cells: ( C ) A representative recording, showing NS1619-induced (30 µM; 5 min) Em hyperpolarization (decrease in IF), which persists even after TNF-α treatment (5 ng/mL; 5 min). TNF-α itself did not alter the Em. As an internal control, we induced Em depolarization with 45 mM K 2 SO 4 . ( D ) Summary of n = 6 individual FLIPR experiments; * p ≤ 0.05. ( E , F ) Em hyperpolarization with NS1619 (30 µM; 24 h) decreases CCL-2 secretion from human pulmonary microvascular endothelial (HULEC5a) and primary human pulmonary artery endothelial (HPAEC) cells when stimulated with 5 ng/mL or 25 ng/mL of TNF-α, respectively, shown by ELISA. ( G ) Em hyperpolarization with NS1619 (30 µM; 24 h) does not change TNF-α-induced (5 ng/mL; 24 h) IL-6 secretion from human pulmonary endothelial cells (HULEC5a), shown by ELISA. Baseline CCL-2 and IL-6 secretion are not affected by NS1619 treatment ( n = 4–11, * p ≤ 0.05). n = number of separate cell passages, which are considered biological replicates.

Article Snippet: Human pulmonary microvascular endothelial cells (HULEC5a; ATCC-CRL-3244) were purchased from ATCC (Manassas, VA, USA) and cultured in an MCDB131 medium (Calsson Labs, Smithfield, UT, USA) supplemented with 10% FBS (Gibco, Waltham, MA, USA), 1% Penicillin/Streptomycin (Gibco, Waltham, MA, USA), 10 mM L-Glutamine (Gibco, Waltham, MA, USA), 10 ng/mL Epidermal Growth Factor (GenScript, Piscataway, NJ, USA), and 1 μg/mL hydrocortisone (Alfa Aesar, Waltham, MA, USA).

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay, Fluorescence, Control

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: MicroRNA-30a as a candidate underlying sex-specific differences in neonatal hyperoxic lung injury: implications for BPD

doi: 10.1152/ajplung.00372.2018

Figure Lengend Snippet: Differential sex-specific expression on miR-30a in vitro. A and B: miR-30a-3p and miR-30a-5p expression in human pulmonary microvascular endothelial cells (HPMECs) exposed to room air (RA) (RA-5% CO2) and 72 h of hyperoxia (95% O2-5% CO2) (n = 3/group). Values are means ± SE. Significant differences between RA and hyperoxia within each sex are indicated by ***P < 0.001. Significant differences between male and female mice in normoxia or hyperoxia are indicated by #P < 0.05 and ##P < 0.01.

Article Snippet: Neonatal human pulmonary microvascular endothelial cells (HPMECs) were purchased from ScienCell (female lot nos: 5016, 10169, 10160, 15900 and 17799; male lot nos.

Techniques: Expressing, In Vitro

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: MicroRNA-30a as a candidate underlying sex-specific differences in neonatal hyperoxic lung injury: implications for BPD

doi: 10.1152/ajplung.00372.2018

Figure Lengend Snippet: Delta-like ligand 4 (Dll4) expression in male and female human neonatal pulmonary microvascular endothelial cells upon exposure to hyperoxia in vitro: DLL4 mRNA (A) and protein (B) expression in male and female neonatal human pulmonary microvascular endothelial cells (HPMECs) exposed to room air (RA) (RA-5% CO2) and 72 h of hyperoxia (95% O2-5% CO2) (n = 3/group). Values are means ± SE. Significant differences between RA and hyperoxia within each sex are indicated by *P < 0.05 and **P < 0.01. Significant differences between male and female mice in normoxia or hyperoxia are indicated by ##P < 0.01. NS, not significant.

Article Snippet: Neonatal human pulmonary microvascular endothelial cells (HPMECs) were purchased from ScienCell (female lot nos: 5016, 10169, 10160, 15900 and 17799; male lot nos.

Techniques: Expressing, In Vitro

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: MicroRNA-30a as a candidate underlying sex-specific differences in neonatal hyperoxic lung injury: implications for BPD

doi: 10.1152/ajplung.00372.2018

Figure Lengend Snippet: Delta-like ligand 4 (Dll4) is an miR-30a-5p target in neonatal human pulmonary microvascular endothelial cells. TargetScan reported miR-30a binding sites 59–66bp in the 3′UTR of DLL4 (A). Increased mir30a-5p expression after mir30a-5p mimic transfection in HPMECs (B). DLL4 mRNA (C) and protein (D) expression after mir30a-5p mimic transfection in HPMECs. DLL4 mRNA (E) and protein (F) expression after mir30a-5p inhibitor transfection in HPMECs. DLL4 protein expression (G) after mir30a-5p mimic transfection with and without target protector in HPMECs. Values are means ± SE from 3 independent experiments (n = 3/group). Significant differences between indicated groups are indicated by *P < 0.05 and **P < 0.01. HPMEC, human pulmonary microvascular endothelial cell; UTR, untranslated region; NC, normal control.

Article Snippet: Neonatal human pulmonary microvascular endothelial cells (HPMECs) were purchased from ScienCell (female lot nos: 5016, 10169, 10160, 15900 and 17799; male lot nos.

Techniques: Binding Assay, Expressing, Transfection, Control

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: MicroRNA-30a as a candidate underlying sex-specific differences in neonatal hyperoxic lung injury: implications for BPD

doi: 10.1152/ajplung.00372.2018

Figure Lengend Snippet: Effect of miR30a-3p overexpression and inhibition on DLL4 expression in HPMECs. Increased mir30a-3p expression after mir30a-3p mimic transfection in HPMECs (A). DLL4 mRNA (B) and protein (C) expression after mir30a-3p mimic transfection in HPMECs. DLL4 mRNA (D) and protein (E) expression after mir30a-3p inhibitor transfection in HPMECs. Values are means ± SE from three independent experiments (n = 3/group). Significant differences between indicated groups are indicated by *P < 0.05 and **P < 0.01. DLL4, delta-like ligand 4; HPMEC, human pulmonary microvascular endothelial cell. NC, normal control.

Article Snippet: Neonatal human pulmonary microvascular endothelial cells (HPMECs) were purchased from ScienCell (female lot nos: 5016, 10169, 10160, 15900 and 17799; male lot nos.

Techniques: Over Expression, Inhibition, Expressing, Transfection, Control

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: MicroRNA-30a as a candidate underlying sex-specific differences in neonatal hyperoxic lung injury: implications for BPD

doi: 10.1152/ajplung.00372.2018

Figure Lengend Snippet: miR30a-5p inhibition under hyperoxic conditions decreases increases Dll4 expression in female HPMECs. DLL4 mRNA (A) and protein (B) expression after inhibition of mir30a-5p in HPMECs upon exposure to hyperoxia. Values are means ± SE from three independent experiments (n = 3/group). Significant differences between indicated groups are indicated by **P < 0.01. DLL4, delta-like ligand 4; HPMEC, human pulmonary microvascular endothelial cell. NC, normal control.

Article Snippet: Neonatal human pulmonary microvascular endothelial cells (HPMECs) were purchased from ScienCell (female lot nos: 5016, 10169, 10160, 15900 and 17799; male lot nos.

Techniques: Inhibition, Expressing, Control

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: MicroRNA-30a as a candidate underlying sex-specific differences in neonatal hyperoxic lung injury: implications for BPD

doi: 10.1152/ajplung.00372.2018

Figure Lengend Snippet: miR30a-5p mimic increases sprouting angiogenesis in female HPMECs. Angiogenic potential was quantified based on the maximum length of sprouts protruding from HPMEC-coated cytodex-3 microcarrier beads suspended in a fibrin gel (n = 3/group). A: representative images from male and female HPMECs subjected to sprouting angiogenesis assay treated with miR30a-5p mimic or negative control. Scale bar represents 200 μm. B: maximum sprouting distance in male and female HPMECs. Values are means ± SE. Significant differences between treatment and control groups is indicated by *P < 0.05. HPMEC, human pulmonary microvascular endothelial cell.

Article Snippet: Neonatal human pulmonary microvascular endothelial cells (HPMECs) were purchased from ScienCell (female lot nos: 5016, 10169, 10160, 15900 and 17799; male lot nos.

Techniques: Angiogenesis Assay, Negative Control, Control