Journal: Hypertension

Article Title: Resolvin E1 Attenuates Pulmonary Hypertension by Suppressing Wnt7a/β-Catenin Signaling

doi: 10.1161/hypertensionaha.121.17809

Figure Lengend Snippet: Figure 1. ChemR23 is downregulated in pulmonary arteries (PAs) from pulmonary hypertension (PH) rodent models and patients with idiopathic pulmonary arterial hypertension (PAH). A, Relative mRNA levels of BLT1 and ChemR23 in cultured mouse pulmonary arterial smooth muscle cell (PASMCs; left) and human PASMCs (right) in response to hypoxia. *P<0.05 vs control (n=6). B, Relative mRNA levels of BLT1 and ChemR23 in PAs from mice subjected to hypoxia and SU5416 (HySu). *P<0.05 vs normoxia (n=6). C, Relative mRNA levels of BLT1 and ChemR23 in PAs from monocrotaline (MCT)- treated rats. *P<0.05 vs vehicle (n=6). D, Left: Representative immunofluorescence images of ChemR23 (red) and α-SMA (green) expression in PAs from HySu-treated mice. Scale bar: 50 μm; (right) Quantification of ChemR23 expression in left. *P<0.05 vs normoxia (n=6). E, Left: Representative immunofluorescence images of ChemR23 (red) and α-SMA (green) expression in PAs from MCT-treated rats. Scale bar: 50 μm; (right) Quantification of ChemR23 expression in left. *P<0.05 vs vehicle (n=6). F, Left: Representative immunofluorescence images of ChemR23 (red) and α-SMA (green) expression in PAs from patients with PAH. Scale bar: 50 μm; (right) Quantification of ChemR23 expression in (left). *P<0.05 vs normal (n=5–6). Data represent the mean±SEM. Statistical significance was evaluated using the Mann- Whitney U test. α-SMA indicates alpha-smooth muscle actin.

Article Snippet: Primary mouse PASMCs and primary mouse pulmonary arterial endothelial cells (PAECs) were isolated and cultured as previously described.22,29 Human PASMCs (cat: PCS100-023) were purchased from the American Type Culture Collection (Manassas, VA).

Techniques: Cell Culture, Control, Immunofluorescence, Expressing, MANN-WHITNEY

Journal: Hypertension

Article Title: Resolvin E1 Attenuates Pulmonary Hypertension by Suppressing Wnt7a/β-Catenin Signaling

doi: 10.1161/hypertensionaha.121.17809

Figure Lengend Snippet: Figure 5. The resolvin E1 (RvE1)/ChemR23 axis suppresses pulmonary arterial smooth muscle cell (PASMC) proliferation in response to hypoxia via Wnt7a/β-catenin signaling. A, Protein levels of Wnt7a, phosphorylated β-catenin(p-β-catenin), and total β-catenin in hypoxia-challenged mouse PASMCs with RvE1 treatment as determined by Western blotting (n=6). B, Knockdown efficiency of Wnt7a siRNA in mouse PASMCs as assessed by real-time polymerase chain reaction. *P<0.05 vs scramble (n=6). C, Effect of Wnt7a knockdown on the proliferation of hypoxia-challenged mouse PASMCs with RvE1 treatment. *P<0.05 vs scramble (n=6). D, Western blot analysis of the effect of Wnt7a knockdown on Wnt7a/β-catenin signaling components in hypoxia-challenged mouse PASMCs with RvE1 treatment after transfection of ChemR23 siRNA (n=6). E, Up, Representative images of immunofluorescence staining of α-SMA (green) and Wnt7a (red) in pulmonary arteries (PAs) from monocrotaline (MCT)-treated rats with RvE1 treatment. Scale bars: 50 μm; (down) Quantification of Wnt7a expression in PAs in up *P<0.05 vs PBS, #P<0.05 vs vehicle (n=6). F, Up: Representative images of immunofluorescence staining of α-SMA (green) and Wnt7a (red) in PAs from HySu-exposed ChemR23F/FSM22cre and ChemR23F/F mice with RvE1 treatment. Scale bars: 50 μm (down). Quantification of Wnt7a expression in PAs in up. *P<0.05 vs RvE1, #P<0.05 vs ChemR23F/F (n=6). Data represent the mean±SEM. Statistical significance was evaluated using the Mann-Whitney U test or Kruskal-Wallis tests followed by Dunn test. α-SMA indicates alpha-smooth muscle actin; and SM22, smooth muscle protein 22.

Article Snippet: Primary mouse PASMCs and primary mouse pulmonary arterial endothelial cells (PAECs) were isolated and cultured as previously described.22,29 Human PASMCs (cat: PCS100-023) were purchased from the American Type Culture Collection (Manassas, VA).

Techniques: Western Blot, Knockdown, Real-time Polymerase Chain Reaction, Transfection, Immunofluorescence, Staining, Expressing, MANN-WHITNEY

Journal: Hypertension

Article Title: Resolvin E1 Attenuates Pulmonary Hypertension by Suppressing Wnt7a/β-Catenin Signaling

doi: 10.1161/hypertensionaha.121.17809

Figure Lengend Snippet: Figure 6. Resolvin E1 (RvE1)/ChemR23 axis inhibits the transcription expression of Wnt7a in hypoxia-challenged mouse vascular smooth muscle cells (VSMCs) through suppressing PKA (protein kinase A)-mediated Egr (early growth response) 2 phosphorylation. A, Up: Gel electrophoresis of PCR-amplified Egr2 binding motif-containing fragments from mouse Wnt7a promoter in hypoxia-exposed mouse pulmonary arterial smooth muscle cell (PASMCs) with or without RvE1 treatment; (down). ChIP analysis of Egr2 binding to the Wnt7a promoter in RvE1-treated mouse PASMCs. *P<0.05 vs PBS, n=6. B, Amino acid sequence alignment of mouse, human, rat, dog, pig, and frog Wnt7a proteins. Ser349 is indicated by an arrowhead, the box indicates the phospho-PKA substrate motif. C, Effect of Egr2 S349A mutant on forskolin-induced Egr2 phosphorylation, detected as the levels of phospho(p)-PKA-substrate (RRXS*/T*) on flag-tagged WT and S349A mutant Egr2 in mouse PASMCs (n=6). D, Effect of Egr2 S349A mutant on RvE1-induced Egr2 phosphorylation, detected as the levels of p-PKA-substrate (RRXS*/T*) on flag-tagged WT and S349A mutant Egr2 in mouse PASMCs (n=6). E, Luciferase reporter assay of RvE1- regulated Wnt7a promoter activity in mouse PASMCs transfected with flag-tagged WT and S349A mutant Egr2 plasmids. *P<0.05 vs vector, $P<0.05 vs WT, #P<0.05 vs PBS (n=6). F, Schematic illustration of ChemR23-mediated inhibition on PASMC proliferation via suppression of Wnt7a-β-catenin signaling in a Gi/PKA/Egr2-dependent manner. Data represent the mean±SEM. Statistical significance was evaluated using Kruskal-Wallis tests followed by Dunn test.

Article Snippet: Primary mouse PASMCs and primary mouse pulmonary arterial endothelial cells (PAECs) were isolated and cultured as previously described.22,29 Human PASMCs (cat: PCS100-023) were purchased from the American Type Culture Collection (Manassas, VA).

Techniques: Expressing, Phospho-proteomics, Nucleic Acid Electrophoresis, Amplification, Binding Assay, Sequencing, Mutagenesis, Luciferase, Reporter Assay, Activity Assay, Transfection, Plasmid Preparation, Inhibition

Journal: The American journal of pathology

Article Title: BDNF/TrkB signaling augments smooth muscle cell proliferation in pulmonary hypertension.

doi: 10.1016/j.ajpath.2012.08.028

Figure Lengend Snippet: Figure 1. Ntrk2 is up-regulated in mouse lungs after 24 hours of hypoxia exposure (pO2 0.01). Total RNA from mice lung homogenates was isolated (n 18 per group), direct labeled, and hybridized to 44K 60mer oligonucleotide microarrays (MWG). A: Volcano plot: log2-fold regulation compared to probability of regulation. Genes with log odds values 5 were considered to be regulated; black spots, exemplary genes. B: Functional annotation of regulated genes was performed according to Gene Ontology (GO). Ntrk2 (TrkB) belongs to MAPK signaling pathway. C: A time course of TrkB expression was performed using real-time PCR of lung homogenates harvested 1, 7, or 21 days after hypoxia exposure (n 7 to 9, each). D: Immunohistochemical staining of TrkB and its ligand, BDNF in mouse lung sections. The arrow indicates positive staining. E and F: Real-time PCR analysis of TrkB and BDNF expression in (E) isolated major pulmonary arteries (n 6), and (F) laser-microdissected intrapulmonary vessels (40 vessels per animal, n 4), from mice exposed to 21 days of hypoxia or normoxia. G: Expression levels of TrkB and BDNF in mouse PASMC as indicated by real-time PCR; lower CT values represent a more abundant transcript level. H: Immunofluorescence staining of TrkB and BDNF in mouse PASMC. The arrows indicate TrkB localization. Inset, negative control staining. *P 0.05.

Article Snippet: To investigate proliferation of unstimulated or BDNF-stimulated human PASMC, cells were permeabilized for 5 minutes with 0.25% Triton X-100 in Tris-buffered saline, incubated with a mouse anti-human Ki-67 antibody (Santa Cruz Biotechnology), followed by incubation with fluorescein isothiocyanate–labeled secondary antibody (Dianova, Hamburg, Germany).

Techniques: Isolation, Labeling, Functional Assay, Expressing, Real-time Polymerase Chain Reaction, Immunohistochemical staining, Staining, Immunofluorescence, Negative Control

Journal: The American journal of pathology

Article Title: BDNF/TrkB signaling augments smooth muscle cell proliferation in pulmonary hypertension.

doi: 10.1016/j.ajpath.2012.08.028

Figure Lengend Snippet: Figure 3. Up-regulation of TrkB and BDNF in human lung tissue from IPAH. A: Colocalization of BDNF and TrkB with SMA as indicated by immunofluorescence staining in donor and IPAH patients. B: Real-time PCR analysis of TrkB and BDNF in laser-microdissected intrapulmonary vessels from donor and IPAH patients. Expression is given in comparison to samples from donor lungs (n 5 per group). C: Real-time PCR analysis of expression levels of TrkB and BDNF in human PASMC; lower CT values represent a more abundant transcript level. D: Immunofluorescence staining of TrkB and BDNF in human PASMC derived from donor and IPAH patients. E: Colocalization of BDNF and TrkB with SMA in PASMC as indicated by immunofluorescence staining, IgG, isotype control staining. *P 0.05.

Article Snippet: To investigate proliferation of unstimulated or BDNF-stimulated human PASMC, cells were permeabilized for 5 minutes with 0.25% Triton X-100 in Tris-buffered saline, incubated with a mouse anti-human Ki-67 antibody (Santa Cruz Biotechnology), followed by incubation with fluorescein isothiocyanate–labeled secondary antibody (Dianova, Hamburg, Germany).

Techniques: Immunofluorescence, Staining, Real-time Polymerase Chain Reaction, Expressing, Comparison, Derivative Assay, Control

Journal: The American journal of pathology

Article Title: BDNF/TrkB signaling augments smooth muscle cell proliferation in pulmonary hypertension.

doi: 10.1016/j.ajpath.2012.08.028

Figure Lengend Snippet: Figure 6. BDNF regulates and activates Egr-1. A: Induction of early growth-response factor 1 (Egr-1) protein in human PASMC as analyzed by Western blotting. B: BDNF-induced Egr-1 protein localization in PASMC as assessed by immunofluorescence analysis. C: Electrophoretic mobility shift assay for Egr-1 induction and DNA–protein interactions from nuclear extracts prepared from BDNF (10 ng/mL)-treated PASMC, incubated with radiolabeled oligonucleotide containing Egr-1 consensus sequence. Incubation with an excess (10-fold) of an unlabeled probe served as a control for specific binding. PDGF (10 ng/mL) treated cells served as a positive control. D: Egr-1 expression in lung parenchyma of donor and IPAH patients. An Egr-1/ mouse (KO-Egr-1) served as negative control. E: Real-time PCR analysis for Egr-1 expression in IPAH patient and donor laser-microdissected intrapulmonary vessels (n 5). n.s., not significant. *P 0.05.

Article Snippet: To investigate proliferation of unstimulated or BDNF-stimulated human PASMC, cells were permeabilized for 5 minutes with 0.25% Triton X-100 in Tris-buffered saline, incubated with a mouse anti-human Ki-67 antibody (Santa Cruz Biotechnology), followed by incubation with fluorescein isothiocyanate–labeled secondary antibody (Dianova, Hamburg, Germany).

Techniques: Western Blot, Immunofluorescence, Electrophoretic Mobility Shift Assay, Incubation, Sequencing, Control, Binding Assay, Positive Control, Expressing, Negative Control, Real-time Polymerase Chain Reaction

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Intracellular iron deficiency in pulmonary arterial smooth muscle cells induces pulmonary arterial hypertension in mice.

doi: 10.1073/pnas.1822010116

Figure Lengend Snippet: Fig. 1. Loss of HAMP responsiveness in PASMCs leads to PAH and right heart failure. (A) Representative images of α-SMA and FPN immunostaining in the lung 6 wk posttamoxifen induction (n = 6 mice per group). Original magnification 40×. (Scale bar: 25 μm.) (B) Representative images of α-SMA (red) and FPN (green) immunostaining in primary mouse PASMCs. Cells were treated with either vehicle or exogenous HAMP peptide (+HAMP). Original magnification 20×. (Scale bar: 100 μm.) (n = 3 mice) (C) Membrane FPN levels measured by flow cytometry in primary mouse PASMCs (n = 3 mice). (D) Representative images of α-SMA immunostaining in the lungs at 6 and 12 wk posttamoxifen induction (n = 6 per group). Original magnification 40×. (Scale bar 25: μm.) (E) Percentage of nonmuscularized (N), partially muscularized (P), and fully muscularized (F) vessels in lungs at 6 wk and 12 wk posttamoxifen induction (n = 5–6 per group). P values shown relative to the respective vessel category in control mice of the respective time point. (F–H) Measurements of right ventricular systolic pressure (RVSP) (n = 5–6), right ventricular ejection fraction (RVEF) (n = 5–12) and right ventricular end systolic volume (RVESV) (n = 5–12). (I) Representative mid- ventricular magnetic resonance images of the heart showing septal bowing at 6 wk post tamoxifen induction (n = 5–12 per group). (J) Ratio of RV to LV+Septum weight (n = 5–6 per group). (K) Representative images of WGA staining in RV (n = 5–6 per group). Original magnification 10×. (Scale bar: 200 μm.) Quantitation of cardiomyocyte area shown in Right. P values for paired comparisons were calculated using Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Mouse PASMCs were treated with mouse HAMP peptide (PLP-4434-s, Peptides International) while human PASMCs were treated with human HAMP peptide (PLP-4392-s, Peptides International) at a final concentration of 0.5 μmol/L for 12–16 h. Ferric citrate (F3388, Sigma-Aldrich) was dissolved in cell growth medium at a final concentration of 200 μmol/L.

Techniques: Immunostaining, Membrane, Flow Cytometry, Control, Staining, Quantitation Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Intracellular iron deficiency in pulmonary arterial smooth muscle cells induces pulmonary arterial hypertension in mice.

doi: 10.1073/pnas.1822010116

Figure Lengend Snippet: Fig. 4. The HAMP/FPN axis operates cell autonomously in PASMCs and is dysregulated by BMPR2 mutations. (A) Representative images of FPN (green) and α-SMA (red) staining (Left) and FPN flow cytometry (Right) in mouse PASMCs. Original magnification 20×. (Scale bar: 100 um.) (B) Representative images of FPN (green) and α-SMA (red) staining (Left) and FPN flow cytometry (Right) in human PASMCs. Cells treated with vehicle or exogenous HAMP peptide (+HAMP) are also shown. Original magnification 20×. (Scale bar: 100 um.) (C–G) Levels of total elemental iron, cellular ferritin, and relative levels of tfr1, dmt1, and et-1 in mouse and human PASMCs treated with scrambled siRNA, hamp siRNA, vehicle, or HAMP peptide (+HAMP). (H) Relative levels of hamp mRNA expression in mouse and human PASMCs with intact BMPR2 or with the R899X mutation. (I) Representative images of FPN flow cytometry in mouse and human PASMCs. R899X cells were treated with vehicle (R899X) or exogenous HAMP peptide (R899X+HAMP). (J) ET-1 levels in supernatants from corresponding samples. (K) Fold induction by BMP6 of hamp mRNA in human and mouse PASMCs with intact BMPR2 or with the R899X mutation. P values are shown relative to R899X cells for the respective BMP6 concentration. (L) ET-1 levels in the supernatants of primary PASMCs derived from three healthy controls (HC1, -2, and -3) and three PAH patients with known BMPR2 mutations (PAH1, -2, and -3). Cells were treated with either vehicle, FAC, or HAMP peptide. P values shown are relative to vehicle-treated cells. (Right) Mean values for all HC vs. all PAH. (E–J and K) P values for paired comparisons were calculated using Student’s t test. (J and L) P values were calculated using ANOVA. Post hoc tests used Bonferroni correction to allow for multiple comparisons. *P < 0.05, **P < 0.01, ***P < 0.001. n = 3 biological replicates.

Article Snippet: Mouse PASMCs were treated with mouse HAMP peptide (PLP-4434-s, Peptides International) while human PASMCs were treated with human HAMP peptide (PLP-4392-s, Peptides International) at a final concentration of 0.5 μmol/L for 12–16 h. Ferric citrate (F3388, Sigma-Aldrich) was dissolved in cell growth medium at a final concentration of 200 μmol/L.

Techniques: Staining, Flow Cytometry, Expressing, Mutagenesis, Concentration Assay, Derivative Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Intracellular iron deficiency in pulmonary arterial smooth muscle cells induces pulmonary arterial hypertension in mice.

doi: 10.1073/pnas.1822010116

Figure Lengend Snippet: Fig. 1. Loss of HAMP responsiveness in PASMCs leads to PAH and right heart failure. (A) Representative images of α-SMA and FPN immunostaining in the lung 6 wk posttamoxifen induction (n = 6 mice per group). Original magnification 40×. (Scale bar: 25 μm.) (B) Representative images of α-SMA (red) and FPN (green) immunostaining in primary mouse PASMCs. Cells were treated with either vehicle or exogenous HAMP peptide (+HAMP). Original magnification 20×. (Scale bar: 100 μm.) (n = 3 mice) (C) Membrane FPN levels measured by flow cytometry in primary mouse PASMCs (n = 3 mice). (D) Representative images of α-SMA immunostaining in the lungs at 6 and 12 wk posttamoxifen induction (n = 6 per group). Original magnification 40×. (Scale bar 25: μm.) (E) Percentage of nonmuscularized (N), partially muscularized (P), and fully muscularized (F) vessels in lungs at 6 wk and 12 wk posttamoxifen induction (n = 5–6 per group). P values shown relative to the respective vessel category in control mice of the respective time point. (F–H) Measurements of right ventricular systolic pressure (RVSP) (n = 5–6), right ventricular ejection fraction (RVEF) (n = 5–12) and right ventricular end systolic volume (RVESV) (n = 5–12). (I) Representative mid- ventricular magnetic resonance images of the heart showing septal bowing at 6 wk post tamoxifen induction (n = 5–12 per group). (J) Ratio of RV to LV+Septum weight (n = 5–6 per group). (K) Representative images of WGA staining in RV (n = 5–6 per group). Original magnification 10×. (Scale bar: 200 μm.) Quantitation of cardiomyocyte area shown in Right. P values for paired comparisons were calculated using Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Mouse PASMCs were treated with mouse HAMP peptide (PLP-4434-s, Peptides International) while human PASMCs were treated with human HAMP peptide (PLP-4392-s, Peptides International) at a final concentration of 0.5 μmol/L for 12–16 h. Ferric citrate (F3388, Sigma-Aldrich) was dissolved in cell growth medium at a final concentration of 200 μmol/L.

Techniques: Immunostaining, Membrane, Flow Cytometry, Control, Staining, Quantitation Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Intracellular iron deficiency in pulmonary arterial smooth muscle cells induces pulmonary arterial hypertension in mice.

doi: 10.1073/pnas.1822010116

Figure Lengend Snippet: Fig. 4. The HAMP/FPN axis operates cell autonomously in PASMCs and is dysregulated by BMPR2 mutations. (A) Representative images of FPN (green) and α-SMA (red) staining (Left) and FPN flow cytometry (Right) in mouse PASMCs. Original magnification 20×. (Scale bar: 100 um.) (B) Representative images of FPN (green) and α-SMA (red) staining (Left) and FPN flow cytometry (Right) in human PASMCs. Cells treated with vehicle or exogenous HAMP peptide (+HAMP) are also shown. Original magnification 20×. (Scale bar: 100 um.) (C–G) Levels of total elemental iron, cellular ferritin, and relative levels of tfr1, dmt1, and et-1 in mouse and human PASMCs treated with scrambled siRNA, hamp siRNA, vehicle, or HAMP peptide (+HAMP). (H) Relative levels of hamp mRNA expression in mouse and human PASMCs with intact BMPR2 or with the R899X mutation. (I) Representative images of FPN flow cytometry in mouse and human PASMCs. R899X cells were treated with vehicle (R899X) or exogenous HAMP peptide (R899X+HAMP). (J) ET-1 levels in supernatants from corresponding samples. (K) Fold induction by BMP6 of hamp mRNA in human and mouse PASMCs with intact BMPR2 or with the R899X mutation. P values are shown relative to R899X cells for the respective BMP6 concentration. (L) ET-1 levels in the supernatants of primary PASMCs derived from three healthy controls (HC1, -2, and -3) and three PAH patients with known BMPR2 mutations (PAH1, -2, and -3). Cells were treated with either vehicle, FAC, or HAMP peptide. P values shown are relative to vehicle-treated cells. (Right) Mean values for all HC vs. all PAH. (E–J and K) P values for paired comparisons were calculated using Student’s t test. (J and L) P values were calculated using ANOVA. Post hoc tests used Bonferroni correction to allow for multiple comparisons. *P < 0.05, **P < 0.01, ***P < 0.001. n = 3 biological replicates.

Article Snippet: Mouse PASMCs were treated with mouse HAMP peptide (PLP-4434-s, Peptides International) while human PASMCs were treated with human HAMP peptide (PLP-4392-s, Peptides International) at a final concentration of 0.5 μmol/L for 12–16 h. Ferric citrate (F3388, Sigma-Aldrich) was dissolved in cell growth medium at a final concentration of 200 μmol/L.

Techniques: Staining, Flow Cytometry, Expressing, Mutagenesis, Concentration Assay, Derivative Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Melk facilitates pulmonary artery smooth muscle cell proliferation and migration in pulmonary hypertension via modulation of YAP/TAZ signaling

doi: 10.3389/fcell.2025.1693346

Figure Lengend Snippet: Maternal Embryonic Leucine Zipper Kinase (MELK) is significantly upregulated in pulmonary artery smooth muscle cells (PASMCs) of pulmonary arterial hypertension (PAH). (A) Heatmap of the GEO dataset GSE144274 showing the top 25 differentially expressed genes (DEGs) in PASMCs from patients with idiopathic pulmonary arterial hypertension (IPAH) and healthy controls. MELK is markedly upregulated in the IPAH group. (B) Volcano plot showing DEGs between PAH and control groups. (C) RT-qPCR analysis showing that human pulmonary artery smooth muscle cells (HPASMCs) treated with PDGF-BB (20 ng/mL, 48 h) exhibited a significant increase in Melk mRNA expression compared with vehicle-treated cells (n = 6). (D) Western blot and quantitative analysis showing that PDGF-BB stimulation (20 ng/mL, 48 h) markedly increased MELK protein expression in HPASMCs (n = 6). (E) Representative immunofluorescence images showing enhanced co-localization of MELK (red) and the SMC marker ACTA2 (green) in the pulmonary arteries of Sugen/hypoxia (Su/H) mice exposed to chronic hypoxia, whereas normoxia control mice displayed minimal MELK expression in the vessel wall. (F) Quantification of MELK fluorescence intensity showing a significant increase in the Su/H + hypoxia group compared with the normoxia group (n = 9). Data are presented as mean ± SD. Two-tailed unpaired Student’s t-test for two-group comparisons (C,D,F) ; FDR-adjusted thresholds for (B) . P < 0.05 was considered statistically significant.

Article Snippet: To address this, we employed a Sugen/hypoxia (Su/H) mouse model of PAH and conducted in vitro studies using human PASMCs to determine the effects of MELK inhibition with OTS167 and MELK overexpression on vascular remodeling, PASMC behavior, and Hippo–YAP/TAZ signaling.

Techniques: Control, Quantitative RT-PCR, Expressing, Western Blot, Immunofluorescence, Marker, Fluorescence, Two Tailed Test

Journal: Frontiers in Cell and Developmental Biology

Article Title: Melk facilitates pulmonary artery smooth muscle cell proliferation and migration in pulmonary hypertension via modulation of YAP/TAZ signaling

doi: 10.3389/fcell.2025.1693346

Figure Lengend Snippet: Effects of MELK inhibition by OTS-167 on PAH and vascular remodeling in Sugen/hypoxia (Su/H) mice. (A) Experimental design. Su/H mice were generated by intraperitoneal (IP) injection of Sugen 5416 (20 mg/kg, once weekly for 3 weeks) and exposure to 10% O 2 for 4 weeks. Starting at the end of week 1, mice received daily IP injections of saline or the MELK inhibitor OTS-167 (5 mg/kg) for 3 weeks. (B) Right ventricular systolic pressure (RVSP) measured by right heart catheterization at week 4 (n = 9 per group). (C) Right ventricular (RV) free wall thickness assessed by echocardiography (Vevo3100) (n = 9 per group). (D) Fulton index [RV/(LV + septum)] determined by postmortem analysis at week 4 (n = 9 per group). (E) Representative hematoxylin and eosin (H&E)–stained lung sections showing pulmonary artery (PA) wall morphology (scale bars = 50 µm). (F) Quantification of PA vascular remodeling, expressed as percentage of vessels with wall thickness >50% of external diameter (n = 9 per group). Data are presented as mean ± SD. Two-way ANOVA followed by Tukey’s post hoc test was used. P < 0.05 was considered statistically significant.

Article Snippet: To address this, we employed a Sugen/hypoxia (Su/H) mouse model of PAH and conducted in vitro studies using human PASMCs to determine the effects of MELK inhibition with OTS167 and MELK overexpression on vascular remodeling, PASMC behavior, and Hippo–YAP/TAZ signaling.

Techniques: Inhibition, Generated, Injection, Saline, Staining

Journal: Hypoxia

Article Title: Hypoxia and the modulation of the actin cytoskeleton – emerging interrelations

doi: 10.2147/HP.S53575

Figure Lengend Snippet: RhoA regulation in different cell types in hypoxia

Article Snippet: Chi et al, 2010 , Rat PASMCs, Rat PAECs , 1.5% O 2 , 24 hrs , Increased RhoA activation.

Techniques: Activation Assay, Activity Assay, Isolation, Expressing, Inhibition