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. 2018 Jun 25;115(31):E7331–E7340. doi: 10.1073/pnas.1805757115

Fig. 2.

Fig. 2.

Kv2 channels impact localization of VAPA and VAPB in HEK cells. (A) VAPA-GFP expressed alone displays uniform localization across the ER. (B) VAPA-GFP expressed with Kv2.1-loopBAD redistributes to Kv2.1-induced ER/PM junctions. (C) VAPA expressed with the ER/PM junction forming protein, JPH4, does not redistribute to junctions. (D) Kv2.2 coexpressed with VAPB-GFP redistributes this VAP to the induced ER/PM junctions. (E) VAPA(K87D/M89D) has a reduced ability to redistribute to Kv2.1-induced ER/PM junctions. (Scale bars: 5 µm.) (F) Bar graph summarizing VAP redistribution by calculating the ratio of fluorescence at ER/PM junctions to that at ER deeper within the cell when junctions are formed using Kv2 and/or JPH4 as indicated. Only Kv2.1 and Kv2.2 increase the ratio of VAP fluorescence. For analysis, a log transformation was used to satisfy the homogeneity of variance condition and a one-way ANOVA was performed, F(8, 36) = 25.699, P = 1.106 × 10−12 with post hoc pairwise Tukey’s tests. *P < 0.0001, significant difference relative to the dsRedER control,. #P < 0.01 significance. Error bars represent SEM. Twenty-five ROIs from five cells were examined in each case.