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GTSE1 is required for profibrotic phenotypic change in <t>fibroblasts</t> 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.
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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.

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