bc ii  (ATCC)

Journal: International Journal of Nanomedicine

Article Title: Extracellular Vesicles Derived from FGF2-Primed Astrocytes Against Mitochondrial and Synaptic Toxicities in Parkinson’s Disease

doi: 10.2147/IJN.S511474

Figure Lengend Snippet: Effects of FGF2-EVs on the mitochondrial function of MPP + pretreated neurons. ( A and B ) Western-blotting of Mfn2 in neurons pretreated with different dose of MPP + , and the quantification. ( C and D ) Western-blotting of Mfn2 in control cells, cells treated with MPP + , MPP + plus CON-EVs, or MPP + plus FGF2-EVs, and the quantification. Notice the up-regulation of Mfn2 in FGF2-EVs treated cells. ( E ) Mito-Tracker staining of cells treated with CON-EVs, MPP + plus CON-EVs, MPP + , MPP + plus FGF2-EVs. The yellow arrows indicate typical mitochondrial morphology in different groups. ( F and G ) Quantification of mitochondrial morphology. ( H and I ) TMRE staining and quantification of mitochondrial membrane potential ( J and K ) DCFH-DA staining and the quantification of ROS levels. Notice the restoration of TMRE staining and ROS levels by FGF2-EVs. N = 3 batches of cells per group in ( A – D ), 6 batches of cells per group in the experiment; ( E – K ). Statistical analyses in K for are performed by One-way ANOVA with Welch Anova multiple comparison test, and others statistical analyses are performed by One-way ANOVA with Tukey’s multiple comparisons test. * P < 0.05. ** P < 0.01. *** P < 0.001. **** P < 0.0001. ns , not significant. Mean ± SEM.

Article Snippet: The cells were collected in the extract solution and further lysed for activity determination using Mitochondrial Complex I (NADH-CoQ Reductase) Activity Assay Kit (E-BC-K834-M, Elabscience, China) and Mitochondrial Complex II Activity Assay Kit (E-BC-K835-M, Elabscience, China) according to the manufacturer’s instructions.

Techniques: Western Blot, Control, Staining, Membrane, Comparison

Journal: International Journal of Nanomedicine

Article Title: Extracellular Vesicles Derived from FGF2-Primed Astrocytes Against Mitochondrial and Synaptic Toxicities in Parkinson’s Disease

doi: 10.2147/IJN.S511474

Figure Lengend Snippet: Effects of NCAM1 silencing on mitochondrial dynamics and function. ( A and B ) Western-blotting of cells treated with CON-EVs, CON-EVs plus MPP + , FGF2-EVs, FGF2-EVs plus MPP + , and the quantification. Notice the higher levels of NCAM1 in cells treated with FGF2-EVs. ( C–G ) Western-blotting of NCAM1, Mfn2, Opa1 and Drp1, and the quantification. si-NCAM1 decreased the expression of Mfn2 and Opa1 while increased the expression of Drp1. ( H – J ) Mito-tracker staining and corresponding measurements of aspect ratio and form factor ( K and L ) TMRE staining and the quantification mitochondrial membrane potential. ( M and N ) DCFH-DA staining and the quantification. si-NCAM1 significantly decreased fluorescent intensity of TMRE while increased that of DCFH-DA. ( O and P ) TUNEL staining and the quantification. si-NCAM1 significantly increased the number of TUNEL-positive cells. N = 3 batches of cells per group in ( A – G ), 6 batches of cells per group ( H – P ). Statistical analyses for N and P were performed using One-way ANOVA with Welch Anova multiple comparison test, and others statistical analyses are performed by One-way ANOVA with Tukey’s multiple comparisons test. * P < 0.05. ** P < 0.01. *** P < 0.001. ns , not significant. Mean ± SEM.

Article Snippet: The cells were collected in the extract solution and further lysed for activity determination using Mitochondrial Complex I (NADH-CoQ Reductase) Activity Assay Kit (E-BC-K834-M, Elabscience, China) and Mitochondrial Complex II Activity Assay Kit (E-BC-K835-M, Elabscience, China) according to the manufacturer’s instructions.

Techniques: Western Blot, Expressing, Staining, Membrane, TUNEL Assay, Comparison

Journal: International Journal of Nanomedicine

Article Title: Extracellular Vesicles Derived from FGF2-Primed Astrocytes Against Mitochondrial and Synaptic Toxicities in Parkinson’s Disease

doi: 10.2147/IJN.S511474

Figure Lengend Snippet: Effects of FGF2-EVs on the expression of NCAM1, mitochondrial and synaptic proteins, and survival of dopamine neurons in PD mice. ( A ) Experimental timeline for EVs treatment, MPTP-induced PD model establishment, and behavioral assessments. CON-EVs / FGF2-EVs were delivered to the lateral ventricle (LV) through continuous cannula. MPTP was administered to establish a PD mouse model. ( B ) Distribution of PKH26-labeled FGF2-EVs in SNc. ( C and D ) Immunofluorescence staining of TH and its quantification in the SNc of mice treated with CON-EVs, CON-EVs plus MPTP, FGF2-EVs, and FGF2-EVs plus MPTP. Notice the increase of the survival of dopaminergic neurons in FGF2-EVs treated PD mice. ( E ) Western-blotting of NCAM1, Mfn2, PSD95 and SYP in SNc region of mice treated with CON-EVs, CON-EVs plus MPTP, FGF2-EVs, and FGF2-EVs plus MPTP. ( F – I ) Quantitative analysis of the expression of NCAM1, Mfn2, PSD95 and SYP in Figure E. Notice the significant up-regulation of NCAM1, Mfn2, PSD95 and SYP by FGF2-EVs treatment. ( J – M ) CatWalk gait analysis of cadence, duration and average speed in mice treated with CON-EVs, CON-EVs plus MPTP, FGF2-EVs, and FGF2-EVs plus MPTP. Statistical analyses for J are performed by Kruskal–Walli’s test, K are performed by One-way ANOVA with Welch Anova multiple comparison test, and others are performed by One-way ANOVA with Tukey’s multiple comparisons test. N = 3 mice per group in ( B – I ), 6 mice per group in ( J – M ). * P < 0.05. ** P < 0.01. *** P < 0.001. ns , not significant. Mean ± SEM.

Article Snippet: The cells were collected in the extract solution and further lysed for activity determination using Mitochondrial Complex I (NADH-CoQ Reductase) Activity Assay Kit (E-BC-K834-M, Elabscience, China) and Mitochondrial Complex II Activity Assay Kit (E-BC-K835-M, Elabscience, China) according to the manufacturer’s instructions.

Techniques: Expressing, Labeling, Immunofluorescence, Staining, Western Blot, Comparison

Journal: International Journal of Nanomedicine

Article Title: Extracellular Vesicles Derived from FGF2-Primed Astrocytes Against Mitochondrial and Synaptic Toxicities in Parkinson’s Disease

doi: 10.2147/IJN.S511474

Figure Lengend Snippet: Graphic summary of the main finding. FGF2-EVs, which are enriched of NCAM1, are effective in alleviating the mitochondrial and synaptic impairment in PD model.

Article Snippet: The cells were collected in the extract solution and further lysed for activity determination using Mitochondrial Complex I (NADH-CoQ Reductase) Activity Assay Kit (E-BC-K834-M, Elabscience, China) and Mitochondrial Complex II Activity Assay Kit (E-BC-K835-M, Elabscience, China) according to the manufacturer’s instructions.

Techniques: