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. 2019 Sep 5;294(43):15743–15758. doi: 10.1074/jbc.RA119.010027

Figure 6.

Figure 6.

TRIP8b with a phosphoablative mutation prevents hyperpolarization of HCN2 gating. A, representative current traces from HEK293 cells stably expressing HCN2 and transiently transfected with GFP and TRIP8b(WT), TRIP8b(S237A), TRIP8b(R234A), or control vector. Whole-cell recordings were performed with cells held at −40 mV in voltage clamp and stepped from −40 to −120 mV. B, quantification of V50; HCN2 + vector: −90.1 ± 1.6 mV (n = 15), HCN2 +TRIP8b(WT): −95.3 ± 1.6 mV (n = 13), HCN2 + TRIP8b(S237A): −88.3 ± 2.2 mV (n = 14), HCN2 + TRIP8b(R234A): −83.9 ± 2.3 mV (n = 15). Independent Student's t tests were performed to compare TRIP8b(WT) to each condition; +TRIP8b(WT) and + vector: p = 0.03, +TRIP8b(WT) and +TRIP8b(S237A): p = 0.02, +TRIP8b(WT) and +TRIP8b(R234A): p < 0.001. C, maximum tail current was divided by the cell capacitance to obtain Ih current amplitude for each condition. HCN2 + vector: 7.2 ± 1.2 pA/pF (n = 15), HCN2 + TRIP8b(WT): 27.3 ± 3.9 pA/pF (n = 13), HCN2 + TRIP8b(S237A): 15.4 ± 4.3 pA/pF (n = 14), HCN2 + TRIP8b(R234A): 18.3 ± 4.0 pA/pF (n = 15). Student's t tests were similarly performed to compare TRIP8b(WT) with each condition; +TRIP8b(WT) and +vector: p < 0.001, +TRIP8b(WT) and +TRIP8b(S237A): p = 0.054, +TRIP8b(WT) and +TRIP8b(R234A): p = 0.1. *, p < 0.05; **, p < 0.01; ***, p < 0.001. All error bars represent mean ± S.E.