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Expression of a long variant of CRACR2A that belongs to the Rab GTPase protein family in endothelial cells.

Wilson LA, McKeown L, Tumova S, Li J, Beech DJ - Biochem. Biophys. Res. Commun. (2014)

Bottom Line: Unexpectedly, short interfering RNA designed to deplete CRACR2A had no effect on CRAC channels in endothelial cells but reduced the abundance of a protein with about twice the mass of CRACR2A.It made a positive contribution to endothelial tube formation.The data suggest that endothelial cells contain a long variant of CRACR2A which is an EF-hand-containing Rab protein that lacks impact on CRAC channels.

View Article: PubMed Central - PubMed

Affiliation: School of Medicine, University of Leeds, Leeds LS2 9JT, UK.

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No effect of CRACR2A siRNA on CRAC channels in endothelial cells. (A) Mean ± s.e.m. paired intracellular Ca2+ data for human umbilical vein endothelial cells (HUVECs) transfected with control siRNA or CRACR2A siRNA (N = 4 each). Thapsigargin (TG, 1 μM) was added in the absence of extracellular Ca2+ before 2 mM Ca2+ was returned to the medium. (B) Summary data for 5 independent experiments of the type illustrated in (A). (C, D) As for (A, B) except using Orai1 siRNA in place of CRACR2A siRNA (n/N = 2/9).
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f0005: No effect of CRACR2A siRNA on CRAC channels in endothelial cells. (A) Mean ± s.e.m. paired intracellular Ca2+ data for human umbilical vein endothelial cells (HUVECs) transfected with control siRNA or CRACR2A siRNA (N = 4 each). Thapsigargin (TG, 1 μM) was added in the absence of extracellular Ca2+ before 2 mM Ca2+ was returned to the medium. (B) Summary data for 5 independent experiments of the type illustrated in (A). (C, D) As for (A, B) except using Orai1 siRNA in place of CRACR2A siRNA (n/N = 2/9).

Mentions: Intracellular Ca2+ was recorded from human umbilical vein endothelial cells (HUVECs) to observe Ca2+ release evoked by thapsigargin (TG) in the absence of extracellular Ca2+ and then CRAC channel-mediated Ca2+ entry as extracellular Ca2+ was added back (Fig. 1A). Unexpectedly, transfection with short interfering RNA (siRNA) targeted to CRACR2A failed to affect Ca2+ release or Ca2+ entry (Fig. 1A and B). In contrast, siRNA targeted to Orai1 suppressed Ca2+ entry but not Ca2+ release, consistent with Orai1-dependent CRAC channels mediating Ca2+ entry (Fig. 1C and D). The data suggest that CRACR2A is unimportant for CRAC channels in these cells.


Expression of a long variant of CRACR2A that belongs to the Rab GTPase protein family in endothelial cells.

Wilson LA, McKeown L, Tumova S, Li J, Beech DJ - Biochem. Biophys. Res. Commun. (2014)

No effect of CRACR2A siRNA on CRAC channels in endothelial cells. (A) Mean ± s.e.m. paired intracellular Ca2+ data for human umbilical vein endothelial cells (HUVECs) transfected with control siRNA or CRACR2A siRNA (N = 4 each). Thapsigargin (TG, 1 μM) was added in the absence of extracellular Ca2+ before 2 mM Ca2+ was returned to the medium. (B) Summary data for 5 independent experiments of the type illustrated in (A). (C, D) As for (A, B) except using Orai1 siRNA in place of CRACR2A siRNA (n/N = 2/9).
© Copyright Policy - CC BY
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4300414&req=5

f0005: No effect of CRACR2A siRNA on CRAC channels in endothelial cells. (A) Mean ± s.e.m. paired intracellular Ca2+ data for human umbilical vein endothelial cells (HUVECs) transfected with control siRNA or CRACR2A siRNA (N = 4 each). Thapsigargin (TG, 1 μM) was added in the absence of extracellular Ca2+ before 2 mM Ca2+ was returned to the medium. (B) Summary data for 5 independent experiments of the type illustrated in (A). (C, D) As for (A, B) except using Orai1 siRNA in place of CRACR2A siRNA (n/N = 2/9).
Mentions: Intracellular Ca2+ was recorded from human umbilical vein endothelial cells (HUVECs) to observe Ca2+ release evoked by thapsigargin (TG) in the absence of extracellular Ca2+ and then CRAC channel-mediated Ca2+ entry as extracellular Ca2+ was added back (Fig. 1A). Unexpectedly, transfection with short interfering RNA (siRNA) targeted to CRACR2A failed to affect Ca2+ release or Ca2+ entry (Fig. 1A and B). In contrast, siRNA targeted to Orai1 suppressed Ca2+ entry but not Ca2+ release, consistent with Orai1-dependent CRAC channels mediating Ca2+ entry (Fig. 1C and D). The data suggest that CRACR2A is unimportant for CRAC channels in these cells.

Bottom Line: Unexpectedly, short interfering RNA designed to deplete CRACR2A had no effect on CRAC channels in endothelial cells but reduced the abundance of a protein with about twice the mass of CRACR2A.It made a positive contribution to endothelial tube formation.The data suggest that endothelial cells contain a long variant of CRACR2A which is an EF-hand-containing Rab protein that lacks impact on CRAC channels.

View Article: PubMed Central - PubMed

Affiliation: School of Medicine, University of Leeds, Leeds LS2 9JT, UK.

Show MeSH
Related in: MedlinePlus