primary human pulmonary arterial adventitial fibroblast cells (paafs  (ScienCell)

Journal: Molecular Therapy

Article Title: GTSE1-driven ZEB1 stabilization promotes pulmonary fibrosis through the epithelial-to-mesenchymal transition

doi: 10.1016/j.ymthe.2024.09.029

Figure Lengend Snippet: GTSE1 is required for profibrotic phenotypic change in fibroblasts and epithelial cells (A) Western blots for analyzing FMT or (B) EMT markers and (C) IF staining for F-actin in cells transfected with shCTRL or sh GTSE1 with or without TGF-β treatment. Small values above each blot reflect the relative intensity of the target proteins normalized to the endogenous control, β-actin. (D and E) Cell migration and fibrotic remodeling of siRNA-transfected cells were assessed with and without TGF-β treatment using (D) wound-healing and (E) gel contraction assays after 24 or 48 h. Results are presented as a fold change of wound closure or gel contraction compared with the original volume. (F) EpCAM (green) was used to identify epithelial adhesion markers co-stained with GTSE1 (red) and α-SMA (violet). All graphs indicate the mean ± SEM ( n = 3/group). The fold change was calculated relative to the control. ∗, ∗∗, ∗∗∗, ∗∗∗∗ indicates p < 0.05, p < 0.01, p < 0.001, p < 0.0001, respectively.

Article Snippet: Next, we performed in vitro experiments using a GTSE1-deficient cell line and primary human pulmonary fibroblast cells purchased from PromoCell (Heidelberg, Germany) to show that GTSE1 enhances EMT signaling in response to injury.

Techniques: Western Blot, Staining, Transfection, Control, Migration

Journal: Molecules (Basel, Switzerland)

Article Title: Ebselen Inhibits the Growth of Lung Cancer Cells via Cell Cycle Arrest and Cell Death Accompanied by Glutathione Depletion.

doi: 10.3390/molecules28186472

Figure Lengend Snippet: Figure 1. Effects of Ebselen on cell viability of lung cancer and normal cells. Exponentially growing cells were incubated with Ebselen at the indicated concentrations for 24 h. Cell growth was evaluated by MTT assays. The chemical figure indicates the structure of Ebselen (2-Phenyl-1,2-benzisoselenazol- 3(2H)-one). Graphs show cell growth of Calu-6 cancer cells (A), A549 cancer cells (B), and primary normal HPF cells (C). * p < 0.05 compared to Ebselen-untreated control cells.

Article Snippet: Primary normal HPF cells were purchased from PromoCell GmbH (C-12360, Heidelberg, Germany) and used between passages five and seven.

Techniques: Incubation, Control

Journal: Molecules (Basel, Switzerland)

Article Title: Ebselen Inhibits the Growth of Lung Cancer Cells via Cell Cycle Arrest and Cell Death Accompanied by Glutathione Depletion.

doi: 10.3390/molecules28186472

Figure Lengend Snippet: Figure 2. Effects of Ebselen on apoptosis in lung cancer and normal cells. Cells in exponential growth phase were incubated with Ebselen at the indicated concentrations for 24 h. Annexin V-FITC and PI staining cells were evaluated with a FAC Star flow cytometer. (A) Representative figures for annexin V-FITC and PI staining cells in lung cancer and normal cells. (B–D) Graphs show proportions of annexin V-positive cells derived from Figure (A) in Calu-6 (B), A549 (C), and HPF cells (D). * p < 0.05 compared to Ebselen-untreated control cells.

Article Snippet: Primary normal HPF cells were purchased from PromoCell GmbH (C-12360, Heidelberg, Germany) and used between passages five and seven.

Techniques: Incubation, Staining, Cytometry, Derivative Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Factors Associated with Heritable Pulmonary Arterial Hypertension Exert Convergent Actions on the miR-130/301-Vascular Matrix Feedback Loop

doi: 10.3390/ijms19082289

Figure Lengend Snippet: Factors genetically associated with PAH are responsive to matrix stiffening. ( A – C ) Pulmonary arterial cells were plated on soft (1 kPa) or stiff (12 kPa) hydrogel for 48 h. Expression level of factors genetically associated with PAH in PAAFs ( A ), PAECs ( B ), and PASMCs ( C ) were analyzed by RT-qPCR. Mean expression in control groups (1 kPa) was assigned a fold change of 1, to which relevant samples were compared. Data are expressed as mean ± SD (* p < 0.05, ** p < 0.01, *** p < 0.001) of at least three independent experiments. Paired samples were compared using a 2-tailed Student’s t -test. ( D ) Schematic of the main results. Red: upregulated genes; Green: downregulated genes; * denote consistent and significant modulation in the three vascular cell types.

Article Snippet: Primary human pulmonary arterial adventitial fibroblast cells (PAAFs) were purchased (Sciencell Research Laboratories, Carlsbad, CA, USA) and grown in FGM cell culture media (Lonza).

Techniques: Expressing, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Factors Associated with Heritable Pulmonary Arterial Hypertension Exert Convergent Actions on the miR-130/301-Vascular Matrix Feedback Loop

doi: 10.3390/ijms19082289

Figure Lengend Snippet: Upstream factors linked to hereditary PAH modulate the PPARγ-APOE-LRP8 axis, which in turn controls pulmonary vascular matrix stiffening. ( A – C ) Following transfection with the indicated siRNAs, transcripts related to the PPARγ-APOE-LRP8-matrix remodeling axis were analyzed by RT-qPCR in PAAFs ( A ), PAECs ( B ), and PASMCs ( C ). Mean expression of a given miRNA in the control group (si-NC) was assigned a fold change of 1, to which corresponding miRNA levels in experimental groups were compared. Data are expressed as mean ± SD (* p < 0.05, ** p < 0.01, *** p < 0.001) of three independent experiments. One-way ANOVA and post-hoc Tukey tests were used for group comparisons. ( D ) Schematic of the main results. In each cell type, the central downstream PPARγ-APOE-LRP8-LOX-collagen axis is drawn. Factors genetically linked to PAH are strategically placed next to the portion of the axis which they regulate; font is also color-coded based on these connections. Blue: genes related to the PPARγ-APOE-LRP8 axis; Red: genes related to fibrillar collagen; Orange: genes related to LOX.

Article Snippet: Primary human pulmonary arterial adventitial fibroblast cells (PAAFs) were purchased (Sciencell Research Laboratories, Carlsbad, CA, USA) and grown in FGM cell culture media (Lonza).

Techniques: Transfection, Quantitative RT-PCR, Expressing