Localization of MAPPER-S relative to STIM1. (A) Cartoon illustrating the spatial organization of the tunneling machinery. (B) Airyscan images of MAPPER–GFP (MAPPER) and mCh-STIM1 following store depletion with CPA in HeLa cells show that they do not colocalize. (C) Violin plots showing the nearest neighbor distance (NND) between the edges of the STIM1 and MAPPER clusters (n = 626–1,585, outliers removed using the ROUT routine, one-way ANOVA). (D) Relative localization of mCh-STIM1 and short MAPPER (MAPPER-S) in control conditions (Ctr) and after store depletion (Tg) at the whole cell level. (E) High magnification images of mCh-STIM1 and MAPPER-S clusters with the corresponding PCC (n = 11, paired t test) and line scan profile measured along the white arrow.