Journal: Cells

Article Title: Transfer of Cardiac Mitochondria Improves the Therapeutic Efficacy of Mesenchymal Stem Cells in a Preclinical Model of Ischemic Heart Disease.

doi: 10.3390/cells12040582

Figure Lengend Snippet: Figure 2. Cardiac mitochondria are internalized by MSCs through dynamin-dependent, clathrin- mediated endocytosis. (A) Representative confocal microscopy pictures of WGA-stained MSCs after 24 h of incubation with MitoTracker Green-labeled cardiac mitochondria at the Mito 3 concentration in the absence or presence of dynasore. Scale bar: 5 µm. (B) Flow cytometry quantification of Mito- Tracker Green-labeled cardiac mitochondria by MSCs following 24 h of exposure in the presence or absence of dynasore (n = 4). (C) Relative Ki67 mRNA levels in cardiac mitochondria-preconditioned MSCs in the presence or absence of dynasore in reference to their respective controls (n = 4). (D) Relative VEGF and HGF mRNA levels in cardiac mitochondria-preconditioned MSCs in the presence or absence of dynasore in reference to their respective controls (n = 4). (E) Relative mRNA levels of CXCL1, CXCL5, CXCL6, IL11, IL33 and LIF in cardiac mitochondria-preconditioned MSCs in the presence or absence of dynasore in reference to their respective controls (n = 4). (F) Relative mRNA levels of MMP1, MMP9 and MMP14 (n =4) and (G) collagenase activity (n = 10) in cardiac mitochondria-preconditioned MSCs in the presence or absence of dynasore in reference to their respective controls. One-way ANOVA with Dunn’s multiple comparisons test in (B–F). One-way ANOVA with Tukey’s multiple comparisons test in (G). * p < 0.05, ** p < 0.01, *** p < 0.001. Each dot represents an independent experiment. Bar graphs represent mean values ± SD.

Article Snippet: To characterize the endocytosis process by which MSCs internalize cardiac mitochondria, human MSCs were exposed to cardiac mitochondria previously labeled with MitoTracker Green FM (40 nM, Invitrogen, Waltham, MA, USA, Cat#M7514) in the presence of the dynamin-dependent, clathrin-mediated endocytosis inhibitor dynasore (50mM, Santa Cruz Biotechnology, Dallas, TX, USA, Cat#sc-202592).

Techniques: Confocal Microscopy, Staining, Incubation, Labeling, Concentration Assay, Flow Cytometry, Activity Assay

Journal: Neuropsychopharmacology

Article Title: Role of Glycine Receptors in Glycine-Induced LTD in Hippocampal CA1 Pyramidal Neurons

doi: 10.1038/npp.2011.86

Figure Lengend Snippet: Gly-LTD was accompanied by rapid internalization of AMPA receptors in CA1 pyramidal neurons. (a) Glycine-induced LTP disappeared when loading recorded cells with the specific SNARE-dependent exocytosis inhibitor, tetanus toxin (0.1 μM, n=6). (b) Suppression of EPSCs was abolished by intracellular loading of the specific dynamin-dependent endocytosis inhibitor, D15 (2 mM, n=6). The persistent changes induced by 1.5 mM glycine (control in c) were borrowed from the data in Figure 1 (c, d) Statistical plots of data showing abolishment of Gly-LTP and LTD by the exocytosis blocker, TeTx, and the endocytosis blocker, D15, respectively. **P<0.01, compared between the indicated groups.

Article Snippet: To investigate whether the Gly-LTD in AMPAR-mediated EPSCs was due to postsynaptic AMPAR internalization, we loaded cells with a specific dynamin-dependent endocytosis inhibitor, D15 (2 mM), which specifically interferes with the binding of dynamin to amphiphysin ( Luscher et al , 1999 ; Morishita et al , 2005 ).

Techniques: Control

Journal: Cells

Article Title: GM101 in Combination with Histone Deacetylase Inhibitor Enhances Anti-Tumor Effects in Desmoplastic Microenvironment

doi: 10.3390/cells10112811

Figure Lengend Snippet: Endocytosis mechanism of Ad in combination with HDACi: ( a ) U343 cells were pretreated with ( a ) dynamin 2 inhibitor (dynasore—40 μM), ( b ) clathrin inhibitor (CPZ—5 μM), or ( c ) caveolin inhibitor (genistein—37 μM) for 45 min. Subsequently, the cells were treated with dAd/GFP, dAd/GFP+SBHA, or dAd/GFP+MS-275. At 24 h post transduction, cells were analyzed for GFP expression by using the IncuCyte ® Live-Cell Analysis System (Sartorius). The data are representatives of three independent experiments performed in triplicate. Representative images are shown. Original magnification: ×40. Bars represent the mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: U343 cells were pretreated with dynamin 2 inhibitor (dynasore—40 μM, Enzo Life Sciences, Farmingdale, NY, USA), clathrin inhibitor (CPZ—5 μM, Enzo Life Sciences), or caveolin inhibitor (genistein—37 μM, Enzo Life Sciences) for 45 min, along with PBS as a negative control.

Techniques: Transduction, Expressing