Fig. 2: Kv3.3 channels are linked to TBK1. | Nature Communications

Fig. 2: Kv3.3 channels are linked to TBK1.

From: Cerebellar Kv3.3 potassium channels activate TANK-binding kinase 1 to regulate trafficking of the cell survival protein Hax-1

Fig. 2

a Western blots showing increased pTBK1 in the cerebellum of G592R Kv3.3 mice compared to that in wild type mice, with no change in total TBK1 levels. b, c Quantification of pTBK1 and total TBK1 (wild type, n = 10; G592R mutant, n = 8 independent experiments, two-tailed unpaired t test for p-TBK1; wild type, n = 3 independent experiments; G592R mutant, n = 3 independent experiments, two-tailed paired t test for total TBK1. Data are mean ± SEM). d, e Controls showing no change in levels of active S6 kinase in mutant animals (wild type, n = 7 independent experiments; G592R mutant, n = 6 independent experiments; data are presented as mean ± SEM, two-tailed unpaired t test). f Blots demonstrating that wild type and G592R Kv3.3 channels co-immunoprecipitate (IP) with TBK1. g Quantification of relative levels of TBK1 co-immunoprecipitated with wild type and G592R Kv3.3 (n = 4 independent experiments; data are mean ± SEM, two-tailed paired t test). h Immunostaining demonstrating colocalization of TBK1(green) with Kv3.3 (red) in CHO cells expressing the channel. Representative images are shown from three independent repeats. Scale bar, 5 μm. i, Fluorescence colocalization analysis by pixel intensity. j Schematic of truncations (TR) of the C-terminus of Kv3.3. k Quantification of the effects of truncations of the Kv3.3 channel C-terminus on the co-immunoprecipitation of TBK1 (n = 4 independent experiments; data are mean ± SEM, one-way ANOVA, Tukey’s multiple comparisons test). Source data are provided as a Source Data file.

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