Figure 1
From: NF-κB-regulated, proinflammatory miRNAs in Alzheimer's disease
Expression of a small family of potentially pathogenic, NF-κB-regulated miRNAs. (A) Light microscopic photograph of human neuronal-glial (HNG) cells in primary culture stained with: antibody to glial fibrillary acidic protein, a glial-specific cytoplasmic marker (green fluorescence; λmax = 556 nm); antibody to βTUBIII, a neuron-specific cytoplasmic marker (red; λmax = 702 nm); and Hoescht 33258 to highlight the morphological features of both glial cell and neuronal cell nuclei (blue; λmax = 461 nm). Note: large nuclear area, relative to both glial or neuronal cytoplasmic area, is indicative of high levels of transcriptional activity [10, 52, 55, 102] (18 days in culture; 20× magnification) [6, 38, 78]. (B) miRNA array cluster analysis and heatmap data from [IL-1β+Aβ42]-induced HNG cells in primary culture; relative induction of homo sapien (hsa)-miRNA-125b, hsa-miRNA-146a, hsa-miRNA-155, hsa-miRNA-9, hsa-miRNA-34a and hsa-miRNA-183 in control (n = 3) and stressed (n = 3) HNG cells by a cocktail of [IL-1β+Aβ42] in combination [38, 52, 78]. Note: twofold to sixfold upregulation in miRNA-125b, miRNA-146a, miRNA-155, miRNA-9 and hsa-miRNA-34a and no significant change in hsa-miRNA-183 (control miRNA in the same sample). (C) [IL-1β+Aβ42]-induced HNG cells in primary culture [38, 78] show significant upregulation of hsa-miRNA-9, hsa-miRNA-125b, hsa-miRNA-146a, and miRNA-155 and quenching of their induction using the polyphenolic resveratrol analog CAY10512 (trans-3,5,4'-trihydroxystilbene). Aβ42+IL-1β has been previously shown to induce NF-κB and proinflammatory miRNA expression in several different human brain primary cell types [38, 53, 72, 78]; other classes of NF-κB inhibitors, including the polyphenolic free radical scavenger curcumin and pyrrolidine dithiocarbamate, have also been shown to significantly quench the upregulation of inducible brain-enriched miRNAs indicating their NF-κB sensitivity [38, 72, 78]. n = 3 to 5; *P <0.01 (analysis of variance), gray bars over white bars (upregulation) or black bars over gray bars (downregulation).