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Figure 4 | Journal of Neuroinflammation

Figure 4

From: The role of the JAK2-STAT3 pathway in pro-inflammatory responses of EMF-stimulated N9 microglial cells

Figure 4

EMF exposure induces expression of p-STAT3 and its DNA-binding activity. Experiments were performed as described above. The protein samples were harvested at the indicated times after EMF exposure in N9 cells. β-Actin antibody was used as a control for equal protein loading. (A) Western blot analysis showing STAT3 activation with antibodies against phospho-STAT3 tyr705 and STAT3; p-STAT3 was labeled with green fluorescence and total STAT3 was labeled with red fluorescence. (B) The amount of p-STAT3 or STAT3/actin ratio was determined by densitometric analysis with software Image J 1.42. (C) STAT3 DNA-binding activity was measured by electrophoretic mobility-shift assays after EMF exposure. No nuclear extract was added to the labeled STAT3 probe (lane 1). EMF sham exposure was used as a control (lane 2). DNA binding was noted at 1, 3, 6, 12 and 24 h after EMF exposure (lane 3-7). P6 inhibited EMF-induced binding activity to STAT3 (lane 8-9). Unlabelled STAT3 was used as competitor DNA at a 200-fold molar excess (Lane 10). A super-shift assay was conducted by pre-incubating of nuclear extracts with anti-STAT3 (lane 11). Results are presented as mean ± S.D. of three independent experiments. Statistical comparisons to control are indicated by * p < 0.05; ** p < 0.01; *** p < 0.001.

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