Figure 6.

Membrane Charge Regulated PIP5K Distribution in the Synapse

(A) CTLs expressing Lifeact-mApple with Kras+8 probe that binds negatively charged areas of the plasma membrane. Upon first contact between CTL and EL4-blue target (t = 0), depletion of probes and recovery with time (min:s). Images are single confocal slices from Video S5A shown as merged and separate channels.

(B) En-face views across 3 μm slice of the synapse as single and merged channels with intensity plot for EGFP-Kras+8.

(C) Pixel intensity (y axis) plots against distance across the synapse/contact site (x axis, μm).

(D) Schematic of the modifications made to the active loop of PIP5Kβ-EGFP.

(E) PIP5K β-EGFP KR_N and PIP5Kβ-EGFP KR_A (both green) during depletion of F-actin (red) at the synapse (100% conjugates, n = 10; all independent experiments). Images are single confocal planes taken from Video S5B (PIP5K β KR_N), and Video S5C (PIP5K β KR_A).

Scale bars = 3 μm.