Effects of depletion of native PIP₂ with CiVSP on R347E and R479E. (A) Normalized currents recorded at 60 mV for R347E in the presence or absence of CiVSP. (B) Activation time constants on cell with (n = 6) or without (n = 5) CiVSP. (C) On-cell G-V curves for R347E controls (black; n = 5) and +CiVSP (red; n = 5) recorded from the typical −100-mV hold (squares) and with a 1-s 60-mV prepulse (circles). Curves show single Boltzmann fits, and mode shift values are reported in Table 3 and plotted in D. (D) R347E mode shift measured on cell with or without CiVSP coexpression compared with WT. (E–H) Repeat of A–D for R479E (n = 3–5 for F and 5–7 for G and H). All data are mean ± SEM (*, P < 0.05; ***, P < 0.001; t test).