Fig. 6

Mn-exposed microglia release a variety of proinflammatory mediators. Multi-plex or single-plex ELISA was utilized to assess the levels of cytokines/chemokines in sampled media from 0 or 100 μM MnCl₂-treated microglia. a Representative image from the Quansys mouse 14-plex array indicating location of noted chemokines and cytokines. b Representative heat maps of Quansys multi-plex ELISA results of media from 0 or 100 μM MnCl₂-treated microglia with dot locations correlating to a. Intensity is represented by no expression (blue) to increasing expression (yellow). Levels of IL-6 (c), CCL2 (d), CCL3 (e), CCL5 (f), and TNF (g) were calculated based on standard curves generated during experiments. Data is represented by mean concentration (pg/mL) ± SEM (Student’s t test; *p < 0.05, **p < 0.01, and ***p < 0.001). NOS2 levels (green) were assessed via immunofluorescence in 0 (h) or 100 μM (i) MnCl₂-treated microglia and quantified by measuring the mean fluorescence intensity per cell (j). Data is represented by mean fluorescence per cell per field ± SEM (Student’s t test; *p < 0.05). Blue = DAPI; scale bar = 10 μm