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Morphological Changes into Neuron-Like Cells and Expression of MAP2 Protein.
Figure 3(d) shows that the expression levels of Map2 mRNA significantly increased with DHA and EPA treatment, reflecting the change in Hes1, Hes6, and NeuroD expression levels in Tuj-1-positive cells.
8C, D), while MAP2 and NFKB1 are detected positive by RT-PCR (Fig.
(b-f) Representative images of dual immunofluorescence indicating coexpression of (b) LMX1-A (green) and FOXA2 (red) in dopaminergic progenitors at day 11; (c) TH (green) and MAP2 (red) in neurons at day 30; (d) TH (green) and DAT (red); (e) TH (red) and GLUR2 (green); and (f) TH (green) and VMAT2 (red) in DA neurons at day 70.
The purity of these cultured cells was identified with specific markers of GFAP and MAP2 for astrocyte and neuron, respectively.
Western Blot Detection of MAP2, PKA, and PPAR-[gamma] Expression.
As recently reported , the electromagnetic waves are able to modulate the cytoskeleton function and to promote the neuronal differentiation of the bone marrow mesenchymal stem cells; in particular, EMFs promote the neuronal differentiation in vitro and the hippocampal neurogenesis in vivo by upregulating the Cav-1 channel activity [25-28], [beta]-III-tubulin, MAP2 , and the brain-derived neurotrophic factor .
(c) Isolated mixed neuronalglial cultures from Sig-1R WT or KO E18 fetuses were seeded at a density of 5 x 104 cells per well and stained with the neuron and astrocyte markers MAP2 (green) and GFAP (red), respectively, on DIV 11.
Subsequent histopathological observations using MAP2 and Iba-1 immunostaining, and analyses of IL-1[beta], TNF[alpha], VEGF, and Bcl-[x.sub.L] expressions demonstrated that RGE promotes neuronal restoration in the injured spinal cord by inhibiting the inflammatory processes and by upregulating the expression of neuroprotective factors (VEGF and Bcl-[x.sub.L]) after SCI.
Primary antibodies included monoclonal antibodies against [beta]-tubulin III, MAP2, tau, GFAP, and [beta]-catenin (Cell Signaling Technology, Danvers, MA, USA).
The differentiated cells were reacted with antibodies against mouse anti-Nestin (1:300, Abcam, Cambridge, MA, USA), mouse antimicrotubule associated protein2 (MAP2, 1:300, Abcam, Cambridge, MA, USA), mouse anti-glial fibrillary acidic protein (GFAP, 1:600, Abcam, Cambridge, MA, USA) and anti-mouse FITC-conjugated IgG antibody (1:500, Abcam, Cambridge, MA, USA).
Results: We demonstrate berberine induces neuronal differentiation accompanying increased neuronal differentiation markers like MAP2, [beta]-III tubulin and NCAM; generated neurons were viable.
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