passages 3 5  (Thermo Fisher)

Journal: iScience

Article Title: TGF-β1 promotes osteogenesis of mesenchymal stem cells via integrin mediated mechanical positive autoregulation

doi: 10.1016/j.isci.2024.110262

Figure Lengend Snippet: Integrin mediated adhesion formation and MSCs osteogenic differentiation correlated with substrate stiffness and TGF-β1 (A) Schematic of the design of cell experiment in vitro for MSCs osteogenesis regulated by TGF-β1 and integrin mediated signaling. (B) Schematic of PEG hydrogel cross-linking process. (C) Immunostaining of Runx2 expression in MSCs w/wo TGF-β1 on hydrogels with different stiffness and the quantification of Runx2 expression (right). Scale bar, 30 μm. (D) Immunostaining of integrin α v β 3 expression in MSCs w/wo TGF-β1 on hydrogels with different stiffness and the quantification of integrin α v β 3 (right). Scale bar, 30 μm. (E) Immunostaining of focal adhesion (FA; paxillin) in MSCs w/wo TGF-β1 on different stiffness hydrogels and the quantification of FA number (upper right) and FA length (lower right). Green, paxillin; red, F-actin; blue, nucleus. Scale bar, 10 μm. (F) Immunostaining of FAK Y397 phosphorylation expression in MSCs w/wo TGF-β1 on different stiffness hydrogels and the quantification of FAK Y397 phosphorylation level (lower). Scale bar, 30 μm ∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05.

Article Snippet: MSCs used in our study have been authenticated by flow cytometry analysis and cell differentiation capacity tests provide by Cyagen Biosciences Inc. 3-5 passage MSCs were seeded onto PEG hydrogel at a density of 1×10 6 cells/mL.

Techniques: In Vitro, Immunostaining, Expressing

Journal: iScience

Article Title: TGF-β1 promotes osteogenesis of mesenchymal stem cells via integrin mediated mechanical positive autoregulation

doi: 10.1016/j.isci.2024.110262

Figure Lengend Snippet: TGF-β1 promoted the integrin-YAP mediated mechanosensing pathway (A) Representative immunofluorescent images of YAP, F-actin, and nuclear lamin A/C-in MSCs w/wo TGF-β1 treatment or on different stiffness hydrogels. Green, YAP; red, F-actin; yellow, lamin A/C; blue, nucleus. Scale bar, 10 μm. (B) Quantification of YAP nuclear/cytoplasm ratio, cytoskeleton anisotropy, and nuclear thickness. (C) Immunostaining of YAP nuclear localization in MSCs after the inhibition of FAK Y397 w/wo TGF-β1 on soft hydrogel and quantification of YAP nuclear/cytoplasm ratio (right). Green, YAP; red, F-actin; blue, nucleus. Scale bar, 30 μm ∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05.

Article Snippet: MSCs used in our study have been authenticated by flow cytometry analysis and cell differentiation capacity tests provide by Cyagen Biosciences Inc. 3-5 passage MSCs were seeded onto PEG hydrogel at a density of 1×10 6 cells/mL.

Techniques: Immunostaining, Inhibition

Journal: iScience

Article Title: TGF-β1 promotes osteogenesis of mesenchymal stem cells via integrin mediated mechanical positive autoregulation

doi: 10.1016/j.isci.2024.110262

Figure Lengend Snippet: TGF-β1 mediated integrin-YAP mechanical positive autoregulation (PAR) (A) Schematic of the proposed pathway of TGF-β1 mediated integrin-YAP mechanical PAR. (B) Immunostaining of FA (paxillin) expression in YAP silenced-MSCs on different stiffness hydrogels. Green, paxillin; blue, nucleus. Scale bar, 10 μm. (C) Quantification of the FA number (left) and FA length (right). (D) Immunofluorescent staining of FA (paxillin) expression in YAP silenced-MSCs with TGF-β1 treatment on the stiff hydrogel. Green, paxillin; blue, nucleus. siNC, control siRNA group. Scale bar, 10 μm. (E) Real-time PCR analysis of FA related genes expression in YAP silenced-MSCs with TGF-β1 treatment on stiff hydrogel, including vinculin, paxillin and talin1. (F) Western blot analysis of FA related protein expression in YAP silenced-MSCs with TGF-β1 treatment on stiff hydrogel, including vinculin, paxillin, and talin1. ∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05.

Article Snippet: MSCs used in our study have been authenticated by flow cytometry analysis and cell differentiation capacity tests provide by Cyagen Biosciences Inc. 3-5 passage MSCs were seeded onto PEG hydrogel at a density of 1×10 6 cells/mL.

Techniques: Immunostaining, Expressing, Staining, Control, Real-time Polymerase Chain Reaction, Western Blot

Journal: iScience

Article Title: TGF-β1 promotes osteogenesis of mesenchymal stem cells via integrin mediated mechanical positive autoregulation

doi: 10.1016/j.isci.2024.110262

Figure Lengend Snippet:

Article Snippet: MSCs used in our study have been authenticated by flow cytometry analysis and cell differentiation capacity tests provide by Cyagen Biosciences Inc. 3-5 passage MSCs were seeded onto PEG hydrogel at a density of 1×10 6 cells/mL.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, RNA Extraction, Real-time Polymerase Chain Reaction, Software