Figure 6

Immunoproteasomes modulate IFNγ-induced inflammation response. BV-2 cells and primary microglia were treated with IFNγ in the absence and presence of immunoproteasome (ONX-0914) and JAK3 (CP-690550, CP) inhibitors, then iNOS and NO levels were measured. (a) Treatment significantly alters iNOS gene expression (F(3,19) = 31.437, p < 0.001). IFNγ (n = 4) significantly increases iNOS gene expression compared to control (n = 5, p < 0.001), whereas IFNγ co-treatment with ONX-0914 (n = 4) results in reduced levels of iNOS gene expression compared to IFNγ alone (p < 0.001). Bars represent mean fold change compared to normalized control. (b) IFNγ and CP-690550 treatment significantly impacts iNOS gene expression (F(3,14) = 16.353, p < 0.001). IFNγ (n = 4) significantly increases iNOS compared to control (n = 5, p < 0.001). IFNγ alone results in significantly higher iNOS gene expression than CP-690550 treatment (n = 5, p = 0.049) and IFNγ + CP-690550 co-treatment (n = 5, p = 0.001). Bars represent mean fold change compared to normalized control. (c) ONX-0914 treatment has a significant effect on NO levels in BV-2 cells (F(3,24) = 68.354, p < 0.001, n = 7). IFNγ treatment increases levels of NO compared to control (p = 0.001). Co-treatment with IFNγ and ONX-0914 results in significantly lower NO production compared to IFNγ alone (p = 0.001). (d) NO levels in primary microglia are significantly different between groups (F(3,8) = 8.717, p = 0.006, n = 3). IFNγ significantly increased NO levels compared to control (p = 0.017), ONX-0914 (p = 0.019) and co-treatment (p = 0.008).