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α-SNAP depleted, resting, or store depleted cells did not show any significant difference in the levels of cell surface Orai1 when compared to controls.
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Blessed with a good harvest, Shakers nonetheless soon found their stores depleted.
Of course Citi is not the only bank out there raising fresh capital to replenish stores depleted by massive write-downs in holdings of mortgage-related securities.
With Ca2+ stores depleted, the residual [Ca2+]i response was spatially uniform (Fig. 3B, gallery b), consistent with Ca2+ entry through VGCCs.
This was further confirmed when subsequent block of VGCCs with 5 μM nicardipine (while keeping Ca2+ stores depleted) completely abolished [Ca2+]i mobilization in response to CCh (Fig. 3B, magenta trace and gallery c).
This could be due to the fact that short inter-pregnancy interval might result in inadequate replenishment of maternal nutrient stores depleted in the previous pregnancy and lead to reduced fetal growth.
Indeed, subsequent inhibition of VGCCs (while keeping Ca2+ stores depleted) completely abolished [Ca2+]i mobilization in response to CCh (Fig. 3B), which also demonstrates that muscarinic cationic channels in SMCs of the guinea-pig ileum are virtually impermeable to Ca2+ (see also [27]).
TIRF images of resting and store-depleted, control, and α-SNAP deficient HEK 293 cells stably expressing CFP-Stim1 and Orai1-YFP.
(n = 3) DOI: http://dx.doi.org/10.7554/eLife.00802.009 We next sought to determine whether α-SNAP, a predominantly cytosolic protein, co-localizes with ER localized Stim1 and/or PM localized Orai1 under resting or store-depleted conditions.
To investigate if the gene expression is functionally relevant, we measured the Ca2+ entry signal occurring when Ca2+ was added back to cells that had been store-depleted by the pharmacological agent thapsigargin.
Notably, over-expression of α-SNAP did not affect total surface Orai1 levels in resting or store-depleted cells or average CFP-Stim1 intensity per cell (data not shown).
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