Journal: Physiological Reports

Article Title: The effect of mitoTEMPO on the development of hypoxia‐induced pulmonary hypertension in male mice

doi: 10.14814/phy2.70804

Figure Lengend Snippet: Effect of mitoTEMPO treatment on HIF‐1α stabilization in vitro. HIF‐1α protein levels were assessed by western blot, and steady‐state mRNA levels for lactate dehydrogenase A ( Ldha ) and pyruvate dehydrogenase kinase 1 ( Pdk1 ) by quantitative real‐time PCR in (a) CMT167 cells, (b) hPASMCs, and (c) mPASMCs, exposed to normoxia (21% O 2 ), severe hypoxia (1% O 2 ), or mild hypoxia (10% O 2 ) for 24 h and treated with triphenylphosphonium (TPP + ) (blue dots) or mitoTEMPO (MT) (red dots). Immunoblots shown are representative of three independent experiments. CMT167: mouse lung carcinoma epithelial cells; hPASMCs: human pulmonary artery smooth muscle cells; mPASMCs: mouse pulmonary artery smooth muscle cells. Densitometric analysis of HIF‐1α bands normalized to β‐Actin. Data are presented as mean ± SD. Statistical comparisons were made using two‐way ANOVA with Tukey's post hoc test ( n = 3 per group). (ns: no signal). Quantitative real‐time PCR data reflect mean ΔCt ± SD ( n = 3 per experimental group).

Article Snippet: Mouse lung carcinoma epithelial (CMT167) cells (10032302, Merck, Germany) and human PASMCs (hPASMCs) (C‐12521, PromoCell, Germany) were purchased.

Techniques: In Vitro, Western Blot, Real-time Polymerase Chain Reaction

Journal: Respiratory Research

Article Title: STAT6 deficiency mitigates the severity of pulmonary arterial hypertension caused by chronic intermittent hypoxia by suppressing Th2-inducing cytokines

doi: 10.1186/s12931-024-03062-z

Figure Lengend Snippet: IL-4 promotes the proliferation of hPASMCs. The concentration of IL-4 ( A ) and IL-13 ( B ) in serum from STAT6 -/- and WT mice after 6 weeks of normoxia or CIH exposure ( n = 6 per group). The concentration of IL-4 ( C ) and IL-13 ( D ) in BALF from STAT6 -/- and WT mice after 6 weeks of normoxia or CIH exposure ( n = 6 per group). ( E ) CCK8 analysis of hPASMCs with IL-4 treatment under concentration of 0, 0.5, 1,10, and 100 ng/ml. ( F ) CCK8 analysis of hPASMCs with IL-13 treatment under concentration of 0, 0.5, 1,10, and 100 ng/ml. ( G ) The percentage of Ki67-positive cells in 10ng/ml IL-13 or IL-4 treated hPASMCs. Data are presented as the mean ± SD. No significant difference is indicated by ns. Significant differences are presented as *( p < 0.05), **( p < 0.01), ***( p < 0.001) and determined by Student’s t-test unless specified. hPASMCs: human pulmonary artery smooth muscle cells; Nor: normoxic; CIH: chronic intermittent hypoxia; WT: wide type; BALF: bronchoalveolar lavage fluid

Article Snippet: hPASMCs were obtained from iCell Bioscience, Inc., Shanghai.hPASMCs were cultured at 37 °C in a humidified incubator with 5% CO 2 using PriMed-iCell-004-LS medium (iCell Bioscience, China).Cells at passages 2–5 were used for further investigation.

Techniques: Concentration Assay

Journal: Respiratory Research

Article Title: STAT6 deficiency mitigates the severity of pulmonary arterial hypertension caused by chronic intermittent hypoxia by suppressing Th2-inducing cytokines

doi: 10.1186/s12931-024-03062-z

Figure Lengend Snippet: IL-4 increased p-STAT6 expression in hPASMCs. ( A ) Western blotting was performed to assess the expression of the STAT6 and p-STAT6 in hPASMCs treated with 10ng/ml IL-4 or IL-13 for 24 h. ( B ) Quantitative analysis of relative expression analysis of STAT6 and p-STAT6 using ImageJ. ( C ) Western blotting was used to detect the expression of STAT6 and p-STAT6 in hPASMCs. ( D - E ) Quantitative analysis of STAT6 and p-STAT6 in hPASMCs using ImageJ. ( F ) Under Nor or IH conditions, hPASMCs were treated with 10 ng/ml IL-4 for 48 h. Cell proliferation was assessed using the CCK8 assay. ( G - H ) Knockdown or overexpression of STAT6 expression in hPASMCs, treat with 10 ng/ml IL-4 and culture for 48 h, then measure hPASMCs proliferation under different conditions using the CCK8 assay. Data are presented as the mean ± SD. No significant difference is indicated by ns. Significant differences are presented as *( p < 0.05), **( p < 0.01), ***( p < 0.001) and determined by Student’s t-test unless specified. Nor normoxia, IH intermittent hypoxia

Article Snippet: hPASMCs were obtained from iCell Bioscience, Inc., Shanghai.hPASMCs were cultured at 37 °C in a humidified incubator with 5% CO 2 using PriMed-iCell-004-LS medium (iCell Bioscience, China).Cells at passages 2–5 were used for further investigation.

Techniques: Expressing, Western Blot, CCK-8 Assay, Knockdown, Over Expression