Fig. 3
From: Key features of the innate immune response is mediated by the immunoproteasome in microglia
Immunoproteasomes mediate IFNγ-dependent cytokine production. BV-2 cells were treated for 24 h, and cytokine levels were measured using a Proteome Profiler assay. (A, B) The heat map displays the mean pixel density from four independent experiments (n = 4 per group). The color scale indicates the range of pixel density values from minimum (blue) to maximum (red). (p < .05) denotes a significant difference between the control and IFNγ treatment groups. ** (p < .05) indicates that co-treatment with ONX-0914 significantly reversed the IFNγ-dependent increase in cytokine levels. For a representative blot image and complete data, please see Supplemental Fig. 3 and the Supplemental Table, respectively. Statistical analysis using one-way ANOVA revealed that ONX-0914 treatment abrogated the IFNγ-dependent increase of Ip-10 ([F(3,11) = 104.4], p < .001). Tukey’s HSD post hoc analysis revealed that IFNγ increased Ip-10 levels compared to control (p < .001), ONX-0914 (p < .001), and ONX-0914 + IFNγ co-treatment (p < .001). In addition, there was a significant difference of Mig protein levels between treatment groups using one-way ANOVA ([F(3,11) = 18.61], p < .001). Tukey’s HSD post hoc analysis revealed that IFNγ treatment resulted in higher Mig protein levels than all other groups (Control p < .001; ONX-0914, p < .001; ONX-0914 + IFNγ, p = .003). MCP-1 levels were significantly different between groups using one-way ANOVA ([F(3,8) = 5.591], p = .02). Tukey’s HSD post hoc analysis revealed that IFNγ treatment increased MCP-1 levels compared to control (p = .035), which was reduced by ONX-0914 co-treatment (p = .029). Rantes protein levels were also different between treatment groups using one-way ANOVA ([F(3,12) = 24.18], p < .001). Tukey’s HSD post hoc analysis revealed that Rantes cytokine levels were significantly higher in the IFNγ treatment group compared to all other groups (Control, p < .001; ONX-0914, p < .001; ONX-0914, +IFNγ, p < .0001). (C) BV-2 β5i KO cells were treated with IFNγ for 24 h, and cytokine levels were measured using a Proteome Profiler assay. Ip-10, Mig, Rantes, and MCP-1 chemokine induction were abrogated in BV-2 β5i KO cells exposed to IFNγ, similar to ONX-0914 treatment.
