Fig. 3

Identification of novel Kv1.3 channel-dependent molecular mechanisms by microglial quantitative proteomics. BV2 microglia were exposed to PBS, LPS (100 ng/mL), ShK-223 (100 nM), or LPS+ShK-223 for 24 h (3 replicates/group) and whole-cell lysates were used for mass-spectrometric analysis. a Volcano-plot: Of 450 proteins differentially expressed across all 4 groups (black dots), 144 proteins were significantly up- or downregulated following LPS exposure (top-right and top-left quadrants; vertical dotted lines represent the upper and lower boundaries of 1.25-fold change threshold while the horizontal dotted line represents p value threshold of 0.05 comparing LPS vs. control groups). Among these, ShK-223 reversed the LPS effect in 21 proteins (highlighted in red). b LPS-upregulated proteins that were reversed by ShK-223 and c LPS-downregulated proteins reversed by ShK-223 are shown. Relative expression represents label-free quantitation data in each treatment condition normalized to the control PBS group. The dotted horizontal line represents the twofold change threshold as compared to the PBS group. d–f Circle plots representing results from GO analyses are shown (gray: protein/gene symbol, red: biological process, green: cellular component, purple: molecular function). Only significantly enriched GO terms (unadjusted p ≤ 0.05 based on enrichment score) are shown (*p < 0.05, **p < 0.01, ***p < 0.005). d GO analysis of 21 LPS-regulated Kv1.3-dependent proteins (n = 21, stringent p ≤ 0.05 threshold comparing LPS vs. LPS+ShK-223). e GO analysis of LPS-upregulated Kv1.3-dependent proteins (n = 26, p ≤ 0.10 threshold comparing LPS vs. LPS+ShK-223). f GO analysis of LPS-downregulated Kv1.3-dependent proteins (n = 40, p ≤ 0.10 threshold comparing LPS vs. LPS+ShK-223)