Fig. 5.

EFR3B(C5S)–TMEM150A interactions facilitate a more rapid recovery of PI(4)P and PI(4,5)P₂ homeostasis than those of other palmitoylated forms of EFR3B. (A–D) HeLa cells were transfected with iRFP–PH(PLCδ), M1R–3×FLAG, and the indicated combination of TMEM150A–GFP and EFR3B–mCherry (EFR3B–mCh) variant (WT or CxS mutant). A time-lapse movie was acquired, recording iRFP fluorescence over time. Shown are kinetics parameters during the post-atropine phase (see Fig. 4): the initial rate of PI(4,5)P₂ recovery (A,B) and maximum rate of PI(4,5)P₂ recovery (C,D). Note that, to improve clarity and facilitate comparisons between effects of either EFR3B mutants alone or co-expressed with TMEM150A, the EFR3B–mCh only data in A are also plotted in B, and all data in C are also plotted in D. (E) Maximum rate of PI(4)P recovery in the presence of the indicated EFR3B and TMEM150A proteins, monitored by the recovery of the PI(4)P biosensor iRFP–P4M to the PM after M1R-mediated depletion. Mean±s.d. are indicated, n=9–21. *P<0.05; **P<0.025; ***P<0.01; ****P<0.001; ns, not significant (two-tailed unpaired t-test in B,D,E for comparisons between two groups; one-way ANOVA with Tukey–Kramer post-hoc test in A,C,E for comparisons between multiple groups).