Fig. 8

Identification of Kv1.3-regulated signaling mechanisms in LPS-induced microglial activation. a, b Canonical pathway analysis of 120 Kv1.3-dependent proteins identified in our proteomic dataset revealed highly represented signaling pathways, two of which are shown here (see Additional file 2: Table S5 for others). Members of this list of 120 proteins are marked with (red circle). Transcription factors are also highlighted (transparent red circle). Arrows indicate directionality of the interaction (upstream vs. downstream). a GABPA, a Kv1.3-dependent transcription factor that was downregulated by LPS, was placed upstream of several Kv1.3-regulated proteins. b MHCI proteins of relevance to our results (TAP1, Tapasin, and EHD1 proteins) were represented in a signaling network that suggested that STAT1 and IRF1 may serve as upstream regulators of these proteins. c Comparison of normalized protein expression of transcription factors identified in our proteomic dataset across treatment groups. d Quantitative RT-PCR data comparing IRF1, IRF7, and NFKB1 mRNA expression across treatment groups (paired t tests were used for these comparisons; six replicates/group). e Phospho-flow cytometric studies of serine (S727) and tyrosine (Y701) STAT1 phosphorylation in BV2 microglia (n = 5/treatment group) at 30-min and 3-h time points. f Immunofluorescence microscopy showing partial co-localization between Kv1.3 (green, detected by ShK-F6CA labeling) and CD14 (red) in LPS-activated BV2 microglia (*p < 0.05, **p < 0.01, ***p < 0.005)