Fig. 4
From: Key features of the innate immune response is mediated by the immunoproteasome in microglia
Altered degradation of IκBα in the absence of immunoproteasome activity. IκBα levels were measured in WT and β5i KO BV-2 cells in the absence and presence of IFNγ. (A) Representative Western blot of IκBα in WT BV-2 and β5i KO BV-2 βcells over a 240 min time course following IFNγ exposure. (B) Quantification of the data represented in (A). Statistical analysis using one-way ANOVA revealed a significant difference between groups ([F(3,28) = 12.75], p < .001). Tukey’s HSD post hoc analysis revealed that IFNγ treatment significantly reduced IkBa levels compared to control (p < .05) after 20 min. IkBa levels were unchanged in BV-2 b5i KO cells treated with IFNγ. *Individual comparisons indicated by asterisks likely used a t-test (n = 4, p < .05, ns = no significance). (C) NF-kB nuclear translocation was measured in BV-2 cells in the presence or absence of IFNγ and/or ONX-0914 by comparing the nuclear to cytoplasmic p65 ratios between groups. An ANOVA revealed a significant difference between groups using one-way ANOVA ([F(3,8) = 11.67], p = .002, n = 3). Tukey’s HSD post hoc analysis revealed that IFNγ treatment increased p65 nuclear to cytoplasmic ratio compared to control (p = .014). Co-treatment with IFNγ and ONX-0914 significantly decreased NF-kB activation compared to IFNγ treatment alone (p = .002), suggesting that immunoproteasome inhibition blocks the IFNγ-dependent activation of NF-kB. (D) Gene expression analysis of cox2 expression by qRT-PCR. Statistical significance between groups (n = 5) was determined using an independent samples t-test (***p < .001, ****p < .0001, ns = no significance).
