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Fig. 8 | Journal of Neuroinflammation

Fig. 8

From: Microglia amplify inflammatory activation of astrocytes in manganese neurotoxicity

Fig. 8

Release of TNF by Mn-activated microglia partly regulates inflammatory microglia-astrocyte crosstalk. Tnf knockdown in microglia was achieved through use of siRNA treatment 48 h prior to MCM experiments. a Successful knockdown of Tnf was assessed in BV2 microglial cells treated with lipopolysaccharide (1 μg/mL) for 24 h after 48 pretreatment with scrambled (Scr) siRNA or Tnf siRNA. b Knockdown of Tnf in primary microglia was assessed via qPCR. c TNF levels in MCM media prior to placement on astrocytes was assessed via single-plex TNF ELISA. d Representative image from the Quansys mouse 14-plex array indicating location of noted chemokines and cytokines. e Representative heat maps of Quansys multi-plex ELISA results of media from 0 or 100 μM MnCl2-treated microglia in combination with Scr siRNA or Tnf siRNA with dot locations correlating to d. Intensity is represented by no expression (blue) to increasing expression (yellow). Levels of inflammatory gene expression in astrocytes treated with MCM of microglia treated with Scr siRNA or Tnf siRNA were determined via qPCR for Tnf (i), Il-1β (j), Il-6 (k), Ccl2 (l), and Ccl5 (m). Expression data is represented by mRNA fold change ± SEM while ELISA data is represented as mean concentration (pg/mL) ± SEM (one-way ANOVA; asterisks above bars indicate significance from control; *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001)

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