primary human pulmonary fibroblast cells  (PromoCell)

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: 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