K_V2.1 channels are not sensitive to depletion of PI(4,5)P₂ by activation of pseudojanin or Dr-VSP. (A) Normalized current traces recorded before (black), during (red), and after (green) application of 10 µM Oxo-M in a cell transiently transfected with K_V2.1 and M₁R. (B) Current traces before (black), after 60 s of application (red), and after washout of 5 µM rapamycin (rapa.) to a cell expressing K_V2.1, pseudojanin-YFP, and LDR-CFP. (C) Normalized K_V2.1-mediated current traces before (black) and 1 s after (red) a 2-s depolarizing pulse to 100 mV in cells coexpressing Dr-VSP. (D) Same as in C but without coexpression of Dr-VSP. (E) Decrease in K_V2.1-mediated steady-state currents at 20 mV after activation of M₁R or Dr-VSP or recruitment of pseudojanin (PJ) to the plasma membrane. Numbers in parentheses indicate numbers of individual experiments. Error bars represent ±SEM.