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. 2014 May 13;5:3771. doi: 10.1038/ncomms4771

Figure 4. IRF8-mediated P2X4R upregulation in microglia depends on IRF5.

Figure 4

Microglial BV2 cells transduced with lentiviral vector encoding IRF5 or control shRNA sequences expressed under a H1 promoter upstream of an EF-1α promoter-IRF8-GFP or GFP expression cassette (shIRF5-IRF8-GFP, shCtrl-IRF8-GFP or shCtrl–GFP). (a) Representative western immunoblot of IRF8 (of four experiments) in whole-cell lysates of BV2 cells transduced with each vector. (b) Real-time PCR analysis of Irf5 mRNA in BV2 cells transduced with each vector (n=6–8; ***P<0.001). (c) Left: representative western immunoblots of IRF5 in BV2 cells transduced with each vector. Right: relative band density ratios of IRF5 (normalized to β-actin) to the shCtrl-GFP-transduced cells (n=4; *P<0.05, ***P<0.001). (d) Real-time PCR analysis of P2rx4 mRNA in BV2 cells transduced with each vector (n=6–8; *P<0.05, **P<0.01). (e) Left: representative western immunoblots of P2X4R in BV2 cells transduced with each vector. Right: relative band density ratios of P2X4R (normalised to β-actin) to the shCtrl-GFP-transduced cells (n=4; *P<0.05, **P<0.01). (f) Real-time PCR analysis of Aif1 mRNA in BV2 cells transduced with each vector (n=6–8; ***P<0.001). Values are the mean±s.e.m. Full-size blots are shown in Supplementary Fig. 11.