Figure 4 | Cell Death & Disease

Figure 4

From: D-ribosylation induces cognitive impairment through RAGE-dependent astrocytic inflammation

Figure 4

RAGE is required for upregulation of p-NF-κB, TNF-α, and GFAP in Rib-treated astrocytoma cells. U251 and U87MG cells were transfected with RAGE siRNA or control siRNA for 24 h and then treated with 20 mM Rib for 2 days. After Rib treatment, the expression levels of p-NF-κB, NF-κB, TNF-α, and GFAP were estimated by western blotting (a). Cells only transfected with control siRNA were used as negative controls and quantification results are shown in panels c, d, g, h, k and l, respectively. For blocking AGE binding to RAGE, U251 and U87MG cells were pre-incubated with anti-RAGE IgG (1 μg/ml). The expression of p-NF-κB, NF-κB, TNF-α, and GFAP of the blocked cells treated with 20 mM Rib for 2 days was analyzed (b). Untreated cells were used as controls. Quantification results were shown in panels e, f, i, j, m and n, respectively. β-Actin was used as a loading control except that NF-κB p65 activation in each sample was expressed as the ratio of phosphor-NF-κB p65 level to total NF-κB p65 level. The control value was set as 1.0. All values are expressed as means±S.E.M. *P<0.05 and **P<0.01 versus control group. #P<0.05 and ##P<0.01 versus Rib group

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