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Fig. 3 | Journal of Neuroinflammation

Fig. 3

From: The overexpression of TDP-43 in astrocytes causes neurodegeneration via a PTP1B-mediated inflammatory response

Fig. 3

PTP1B inhibition and absorption of proinflammatory cytokines mitigate neuronal toxicity caused by TDP-43 overexpression in astrocytes. a, b Neuron-astrocyte coculture. b TDP-43-Gfp-transfected astrocytes were treated with a PTP1B inhibitor (PTP1Bi, 5 μM) or Ptp1b siRNA (50 nM) for 24 h and then were cocultured with primary cortical neurons in transwell culture inserts. Neuronal viability was measured using a CCK-8 assay after a coculture period of 36 or 48 h. Data are presented as the mean ± SD. *p < 0.05; ***p < 0.001 (one-way ANOVA). ce ACM-treated neuron culture. d TDP-43-Gfp-transfected astrocytes were treated with a PTP1B inhibitor (PTP1Bi, 5 μM) or Ptp1b siRNA (50 nM) for 24 h, and then the ACM were harvested. Primary cortical neurons were stimulated with GFP ACM, TDP-43 ACM or TDP-43 + PTP1Bi ACM for 5 days, and then a CCK-8 assay was performed. Data are presented as the mean ± SD. **p < 0.005; ***p < 0.001 (one-way ANOVA). e At the end of neuron-ACM coculture, primary cortical neurons were stained with CMFDA (green). Then, CMFDA-positive neurons were counted under a fluorescence microscope. Data are presented as the mean ± SD of 3. **p < 0.005 (one-way ANOVA). Scale bars, 20 μm. fh Primary cortical neurons stimulated with TDP-43 ACM were treated with IL-1β antibody (50 ng/ml), IL-6 antibody (50 ng/ml) and TNF-α antibody (10 ng/ml) for 5 days and then were subjected to a CCK-8 assay or to CMFDA staining. f CCK-8 assays were performed to assess the viability of primary cortical neurons. Data are presented as the mean ± SD. *p < 0.05; **p < 0.005 (one-way ANOVA). g CCK-8 assays were performed to assess the viability of primary cortical neurons stimulated with TDP-43-transfected ACM and treated with a control IgG antibody (50 ng/ml) for 5 days. Data are presented as the mean ± SD. N.S. not significant (Student’s t test). h CMFDA-positive neurons were counted under a fluorescence microscope. Data are presented as the mean ± SD. **p < 0.005; N.S. not significant (one-way ANOVA). Scale bars, 20 μm

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