Journal: Journal of Nanobiotechnology

Article Title: Exosomes derived from human adipose mesenchymal stem cells ameliorate hepatic fibrosis by inhibiting PI3K/Akt/mTOR pathway and remodeling choline metabolism

doi: 10.1186/s12951-023-01788-4

Figure Lengend Snippet: hADMSCs-Exo inhibits hepatic stellate cell proliferation by impeding cell cycle progression and inducing apoptosis. A A schematic representation of the experimental design. B In vitro cell viabilities of activated LX-2 cells incubated with hADMSCs-Exo at the indicated concentration in the presence of TGF-β1 (10 ng/ml) for 24 h, 48 h or 72 h, (n = 5). C IC50 were analysed in activated LX-2 cells exposed to hADMSCs-Exo for 24 h. D Edu assay showed that HSCs proliferation was suppressed by hADMSCs-Exo in a concentration-dependent manner. EdU% is used as an approximation for proliferation rate. E Cell cycle analysis showed an increase in the sub-G1 subpopulation and cell cycle arrest after hADMSCs-Exo. F The quantitative analysis of the apoptosis was performed using the Annexin-FITC staining based flow cytometry. Exo, exosome. Data are presented as means with SEM (n = 3 independent experiments). ns, not significant, *p < 0.05, **p < 0.01 and ***p < 0.001

Article Snippet: Cell Cycle Analysis Kit (KeyGENEBioTECH, KGA511, China) was used for cell cycle analysis and the Annexin V-FITC & PI Cell Apoptosis Analysis Kit (BD, 550911, USA) was used for apoptosis assays.

Techniques: In Vitro, Incubation, Concentration Assay, EdU Assay, Cell Cycle Assay, Staining, Flow Cytometry

Journal: Journal of Nanobiotechnology

Article Title: Exosomes derived from human adipose mesenchymal stem cells ameliorate hepatic fibrosis by inhibiting PI3K/Akt/mTOR pathway and remodeling choline metabolism

doi: 10.1186/s12951-023-01788-4

Figure Lengend Snippet: hADMSCs-Exo treatment improves liver function and regeneration, reduces liver inflammation and apoptosis. A The Hyp and MDA levels of normal mice (Sham), fbrotic mouse model and the mice injected with PBS, hADMSCs or hADMSCs-Exo were measured with corresponding test kit B Serum levels of AST, ALT, ALP in different groups. C The relative inflammatory gene expression for IL-1β, IL-6, IL-10 and TNF-α. D Immunohistochemical staining was performed to detect the protein expressions of Ki67, HNF-4α and caspase 3 in the injured liver of mice treated with hADMSCs and hADMSCs-Exo. Scale bar, 40 μm. AST, aspartate aminotransferase, ALT, alanine aminotransferase, ALP, alkaline phosphatease. LFG, liver fibrosis group, REG, regression group. Data are expressed as mean ± SEM (n = 4), ns, not significant, *p < 0.05, **p < 0.01 and ***p < 0.001

Article Snippet: Cell Cycle Analysis Kit (KeyGENEBioTECH, KGA511, China) was used for cell cycle analysis and the Annexin V-FITC & PI Cell Apoptosis Analysis Kit (BD, 550911, USA) was used for apoptosis assays.

Techniques: Injection, Expressing, Immunohistochemical staining, Staining

Journal: Journal of Cancer

Article Title: Macrophage-derived SPARC Attenuates M2-mediated Pro-tumour Phenotypes

doi: 10.7150/jca.39651

Figure Lengend Snippet: Effects of SPARC overexpression on M2-mediated anti-apoptosis ability. After treated with 5-fluorouracil at IC50 (1 µg/mL for BGC-823, SGC-7901, MKN-45 and 30 µg/mL for GES-1) for 48 hours, more apoptotic cells were observed by flow apoptosis detection in all four SPARC overexpression groups.

Article Snippet: Cell apoptosis was assessed with the Annexin V-FITC/PI Apoptosis Analysis Kit (Becton, Dickinson, New Jersey, USA).

Techniques: Over Expression

Journal: Antioxidants

Article Title: Magnolia officinalis Bark Extract Prevents Enterocyte Death in a Colitis Mouse Model by Inhibiting ROS-Mediated Necroptosis

doi: 10.3390/antiox11122435

Figure Lengend Snippet: MBE efficiently blocks SZ-induced necroptosis in both InEpCs and HT29 cells. ( A ) Effects of MBE on the viability of InEpC cells treated with the indicated extract concentration for 24 h and 48 h. ( B ) InEpC cells were pretreated with the indicated concentrations of MBE for 30 min before treatment with SZ (Smac mimetic (20 µM) and z−VAD (20 µM)) for 24 h. annexin V/PI was analyzed using flow cytometry. ( C ) The values represent the sum of the percentage of cells in A (PI+ annexin V+; necroptosis). ( D ) Levels of pRIP1, RIP1, pRIP3, RIP3, pMLKL, and MLKL were analyzed using immunoblotting. Tubulin was used as a loading control. ( E ) HT29 cells were treated with the indicated concentrations of MBE for 24 h and 48 h. ( F ) HT29 cells were pretreated with the indicated concentrations of MBE for 30 min before treatment with SZ (Smac mimetic (100 nM) and z−VAD (20 µM)) for 12 h. The expression of annexin V in HT29 cells was induced by SZ and analyzed using flow cytometry. ( G ) The values represent the sum of the percentage of cells in E (PI+ annexin V+; necroptosis). ( H ) Levels of pRIP1, RIP1, pRIP3, RIP3, pMLKL, and MLKL were analyzed using western blotting. Tubulin was used as a loading control. ** p < 0.01, *** p < 0.001, and **** p < 0.0001 (data were analyzed using an ANOVA).

Article Snippet: Cell death was assessed using an Annexin V-FITC Analysis Kit (Cat# BD556547, BD, Franklin Lakes, NJ, USA).

Techniques: Concentration Assay, Flow Cytometry, Western Blot, Expressing

Journal: Antioxidants

Article Title: Magnolia officinalis Bark Extract Prevents Enterocyte Death in a Colitis Mouse Model by Inhibiting ROS-Mediated Necroptosis

doi: 10.3390/antiox11122435

Figure Lengend Snippet: MN and HK inhibit SZ-induced expression of necroptosis proteins in HT29 cells. ( A and D ) HT29 cells were pretreated with MN and HK for 30 min before the treatment with SZ (Smac mimetic (100 nM) and z-VAD (20 µM)) for 12 h. ( B ) The values represent the sum of the percentage of cells in A (PI+ annexin V+; necroptosis). ( C and F ) Protein levels of pRIP1, RIP1, pRIP3, RIP3, pMLKL, and MLKL were analyzed using western blotting. Tubulin was used as a loading control. ( E ) The values represent the sum of the percentage of cells in D (PI+ annexin V+; necroptosis). **** p < 0.0001 (data were analyzed using an ANOVA).

Article Snippet: Cell death was assessed using an Annexin V-FITC Analysis Kit (Cat# BD556547, BD, Franklin Lakes, NJ, USA).

Techniques: Expressing, Western Blot

Journal: Biomolecules

Article Title: Effects of the Leptin-Mediated MAPK/ERK Signaling Pathway on Collagen II Expression in Knee Cartilage of Newborn Male Mice from Obese Maternal Offspring

doi: 10.3390/biom12030477

Figure Lengend Snippet: Expression of key cartilage-related factors under different doses of leptin in vitro. ( A ) Toluidine blue staining of chondrocytes (×200 magnification). ( B ) Representative collagen II immunofluorescent staining (×200 magnification). ( C ) Detection of apoptosis by flow cytometry ( n = 3, chondrocytes obtained from three separate repeated experiments). ( D ) Statistical analysis of apoptosis rate. ( E ) Representative western blot bands of the expression of p-ERK in the nucleus and collagen II, MMP13, RUNX2, MMP1, Leptin Receptor, SOX9, and TIMP1 in the cell cytoplasm induced by leptin in vitro ( n = 3, chondrocytes obtained from three separate repeated experiments). ( F ) Statistical analysis of the relative protein levels. ( G ) Relative mRNA expression of cartilage-related factors under different doses of leptin treatment for 24 h in vitro ( n = 4, chondrocytes obtained from four separate repeated experiments). Results are expressed as mean ± SD. ns not significant, * p < 0.05, *** p < 0.001.

Article Snippet: Annexin V-FITC/propidium iodide (PI) Cell Apoptosis Analysis Kit (BD, 556547, San Jose, CA, USA) was used for apoptosis assays with a flow cytometer (BD FACSCelesta, San Jose, CA, USA).

Techniques: Expressing, In Vitro, Staining, Flow Cytometry, Western Blot