Journal: Cellular and Molecular Life Sciences

Article Title: eEF2 improves dense connective tissue repair and healing outcome by regulating cellular death, autophagy, apoptosis, proliferation and migration

doi: 10.1007/s00018-023-04776-x

Figure Lengend Snippet: Prognostic biomarker selection and verification in injured Achilles tendon tissues. Association of eEF2 expression with a ATRS and, b average heal rise, n = 40; c eEF2 expression between good outcome (GO, n = 20) and poor outcome (PO, n = 20) patients, data presented as mean \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\pm$$\end{document} ± SD, *** p < 0.001; Association among FBN2 and d ATRS and, e average heal rise, n = 40; f FBN2 levels among good outcome (GO, n = 20) and poor outcome (PO, n = 20) patients, data presented as mean \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\pm$$\end{document} ± SD, *** p < 0.001; g AUC for eEF2 to report its predictive significance, n = 40; h Semi-quantitative analysis of eEF2 western blot analysis among good outcome (GO, n = 9) and poor outcome (PO, n = 9) patient samples, data presented as mean \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\pm$$\end{document} ± SD, ** p < 0.01, n = 9 per group; i Positive association between eEF2 and 1-year healing outcome based on western blot analysis, n = 18; j Western blot image of eEF2 and beta-actin (β-actin) in good (G) and poor (P) outcome patients

Article Snippet: Monoclonal antibody to eEF2 (#MA5-42937; Thermo Fisher Scientific) and a secondary antibody, goat anti-rabbit (#31460; Thermo Fisher Scientific, 1:250) was used for immunolocalization.

Techniques: Biomarker Discovery, Selection, Expressing, Western Blot

Journal: Cellular and Molecular Life Sciences

Article Title: eEF2 improves dense connective tissue repair and healing outcome by regulating cellular death, autophagy, apoptosis, proliferation and migration

doi: 10.1007/s00018-023-04776-x

Figure Lengend Snippet: eEF2 expression and localization in tissue biopsies and association with Col1a1. Immunohistochemistry of the eEF2 in a good outcome (GO) and, b poor outcome (PO) patients, Scale bars = 100 µm; c semi-quantitative analysis of eEF2 expression in the good outcome (GO) and poor outcome (PO) patient samples presented as integrated optical density (IOD), data reported as mean \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\pm$$\end{document} ± SD, *** p < 0.001, n = 9 per group. eEF2 immunofluorescence signal in a d good outcome (GO) and, e poor outcome (PO) patient sample. Scale bars = 100 µm; f Western blot analysis and g semi-quantitative analysis of Col1a1 expression in good and poor outcome patients. Signal intensity was used for quantitative analysis, and the intensity of the house-keeping gene (beta-actin) used for normalization; h association among eEF2 and Col1a1 expression, n = 18

Article Snippet: Monoclonal antibody to eEF2 (#MA5-42937; Thermo Fisher Scientific) and a secondary antibody, goat anti-rabbit (#31460; Thermo Fisher Scientific, 1:250) was used for immunolocalization.

Techniques: Expressing, Immunohistochemistry, Immunofluorescence, Western Blot

Journal: Cellular and Molecular Life Sciences

Article Title: eEF2 improves dense connective tissue repair and healing outcome by regulating cellular death, autophagy, apoptosis, proliferation and migration

doi: 10.1007/s00018-023-04776-x

Figure Lengend Snippet: eEF2 enhances healing processes in TNF-induced inflammatory fibroblast models. a – f eEF2 affects Col1a1 expression through autophagy in human fibroblasts. a , b Representative Western blot images and semi-quantitative analysis of LC3-II and LC3-II/LC3-I ratio, c – f representative Western blot ( c ) and confocal images ( e ) along with semi-quantitative analysis of Col1a1 ( d , f ) in cells treated with normal medium, starved medium for autophagy, si-eEF2 and 3-MA incubation based on autophagy, TNF was used afterward to create an inflammatory model; signal intensity ( a – d ) and fluorescent green intensity ( e , f ) were used for semi-quantitative analysis; g – i representative images captured by fluorescent microscope demonstrated the cell death and apoptosis when treated with normal condition, TNF and si-eEF2 and TNF: si-eEF2 increases the ratio of dead/live cells among primary fibroblast and fibroblast cell line; si-eEF2 positively associate with cell apoptosis among human fibroblasts h , j ; the ratio of dead/live cells was reported by percentage and the apoptotic level of cells was presented by fluorescent green intensity. Data reported as mean \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\pm$$\end{document} ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, scale bars = 100 µm, three replicates were used for quantitative analysis

Article Snippet: Monoclonal antibody to eEF2 (#MA5-42937; Thermo Fisher Scientific) and a secondary antibody, goat anti-rabbit (#31460; Thermo Fisher Scientific, 1:250) was used for immunolocalization.

Techniques: Expressing, Western Blot, Incubation, Microscopy

Journal: Cellular and Molecular Life Sciences

Article Title: eEF2 improves dense connective tissue repair and healing outcome by regulating cellular death, autophagy, apoptosis, proliferation and migration

doi: 10.1007/s00018-023-04776-x

Figure Lengend Snippet: eEF2 enhances fibroblast proliferative processes. a , b Representative confocal images and semi-quantitative analysis of Col1a1 in primary fibroblasts and fibroblast cell line, with and without si-eEF2; c , d representative immunofluorescence images and number of proliferating human fibroblasts, with and without si-eEF2; e , f representative immunofluorescence images and ratio of dead/live fibroblasts, with and without si-eEF2; g , h representative immunofluorescence images and number of apoptotic cells from primary fibroblasts and fibroblast cell line, with and without si-eEF2; i – k representative images and quantitative analysis of cell migration rate assessed at 0 and 24 h in primary fibroblast and cell line, with and without si-eEF2. Data reported as mean \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\pm$$\end{document} ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, scale bars = 100 µm, n = 3 replicates

Article Snippet: Monoclonal antibody to eEF2 (#MA5-42937; Thermo Fisher Scientific) and a secondary antibody, goat anti-rabbit (#31460; Thermo Fisher Scientific, 1:250) was used for immunolocalization.

Techniques: Immunofluorescence, Migration

Journal: Cellular and Molecular Life Sciences

Article Title: eEF2 improves dense connective tissue repair and healing outcome by regulating cellular death, autophagy, apoptosis, proliferation and migration

doi: 10.1007/s00018-023-04776-x

Figure Lengend Snippet: Potential mechanisms and eEF2 mode of action during inflammation and proliferation phases of dense connective tissue repair

Article Snippet: Monoclonal antibody to eEF2 (#MA5-42937; Thermo Fisher Scientific) and a secondary antibody, goat anti-rabbit (#31460; Thermo Fisher Scientific, 1:250) was used for immunolocalization.

Techniques: