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Fig. 7 | Molecular Neurodegeneration

Fig. 7

From: Immunotherapy targeting toll-like receptor 2 alleviates neurodegeneration in models of synucleinopathy by modulating α-synuclein transmission and neuroinflammation

Fig. 7

Astrocyte responses by TLR2 mediated neuron-to-astrocyte α-synuclein transmission. a Overview diagram. Donor neuronal cells (V1S), expressing α-synuclein-conjugated with N-terminus of venus were plated in trans-well insert and the recipient human primary astrocytes were plated onto the cover slips in the bottom well. Only astrocytes were treated with either lentiviral vectors or antibodies. Images were taken from astrocytes after a 3-days co-culture. b, c Representative confocal images for N-terminus of venus (Upper panel) and IL-6 (Lower panel) in recipient astrocytes. The fluorescence intensity of N-term venus and IL-6 were analyzed in randomly chosen area. b V1S and astrocytes were co-cultured in the presence of either LV-control/sh.control, LV-TLR2, or LV-sh.TLR2 (n = 3). c V1S and astrocytes were co-cultured in the presence of either IgG (5 μg/ml) or T2.5 (5 μg/ml) (n = 3). d The kinetics of astroglial α-synuclein internalization in the presence of antibodies. Human primary astrocytes were incubated with αSCM for indicated hours in the presence of either IgG (5 μg/ml) or T2.5 (5 μg/ml). The kinetics was analyzed by immunolabeling assay (n = 3). e–h Quantitative analysis of the cytokine/chemokine gene expressions in astrocytes. The cells were incubated with either LZCN or αSCM for 24 h in the presence of indicated antibodies. The expressions of IL-1β (e), IL-6 (f), CCL5 (g), and CX3CL1 (h) were normalized to the levels of β-actin (n = 4). Data are mean ± SEM. *p < 0.05, **p < 0.01, and ***p < 0.001; one way ANOVA for all analysis except (d) (unpaired t test). Scale bar, 20 μm

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