Fig. 8
From: Glial-neuronal signaling mechanisms underlying the neuroinflammatory effects of manganese
Selective knockdown of the astrocyte-specific chemokine, Ccl2, with siRNA in mixed glia does not inhibit Mn-induced inflammatory gene expression but prevents neuronal cell death. a Ccl2 KD does not inhibit mRNA expression of inflammatory genes in Mn-exposed mixed glia compared to mixed glia treated with Mn in the presence of control siRNA. b CCL2 protein levels in GCM are reduced in Ccl2 KD mixed glial cultures. c Viability of N2A cells is preserved following treatment with Mn-GCM from Ccl2 KD mixed glia compared to those treated with control siRNA Mn-GCM. d (left two panels) CCL2 is released into medium in both mixed glia (GCM) and pure astrocytes (ACM) following treatment with MnCl2. d (right two panels) Pharmacologic inhibition of NF-κB with Bay-11 in both mixed glia and pure astrocyte cultures inhibits release of CCL2, whereas CCL2 levels are statistically increased compared to control in both Mn-GCM and Mn-ACM treated with vehicle control (DMSO). e C3 KD does not inhibit mRNA expression of inflammatory genes in Mn-exposed mixed glia compared to mixed glia treated with Mn in the presence of control siRNA. f N2A viability is decreased in cells exposed to C3 KD Mn-GCM compared to those treated with siRNA control Mn-GCM. One-way ANOVA analyses performed for experiments comparing three or more treatment groups and t test in those comparing two treatment groups. Data depicted as ± S.E.M. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 (≥ 4 per treatment group; across ≥ 3 independent experiments)