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Fig. 1. STIM1 oligomerization upon maximal ER Ca²⁺ depletion correlated with ER Ca²⁺ depletion, while subplasmalemmal STIM1 clustering was delayed and followed the decay of cytosolic Ca²⁺ levels. (A-D) Representative tracings of the effect of 100 µM histamine on [Ca²⁺]_ER (A), STIM1 oligomerization (B), subplasmalemmal STIM1 clustering (C) and [Ca²⁺]_cyto (D) in EGTA-containing solution followed by re-addition of 2 mM Ca²⁺ in the absence of histamine. (E) Respective statistical evaluation of the experiments displayed in A (n=11), B (n=7), C (n=12) and D (n=12). (F) Schematic illustration of STIM1 activation by ER Ca²⁺ depletion. (G) Images of the subcellular distribution of YFP-STIM1 under conditions of high ER Ca²⁺ levels in resting cells (left images) and low ER Ca²⁺ levels upon cell stimulation with histamine in EGTA (right images) at the middle z-plane (upper images) and at a TIRF-like bottom plane (lower images). (H) Normalized tracings of [Ca²⁺]_ER and [Ca²⁺]_cyto were inverted and plotted with the respective normalized traces for STIM1 oligomerization and STIM1 clustering (_max=100%).^*P<0.05 versus the respective compared data set.

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