1. (red) & endothelium (green) PMCA4 (green) & DAPI (blue)
PMCA4 & DAPI
PMCA4WT
PMCA4KO B. A. C. D.
Aorta
i) VSM
PMCA4 (red) & VSM
(green)
50µM
ii) Endothelium
(red) & endothelium (green)
100 µm
Isolated mesenteric VSM cells
PMCA4 (green) & DAPI (blue) i)
ii)
25 µm
Figure 1: PMCA4 protein is absent from vascular cells and tissues of PMCA4KO mice, but expression of other calcium handling proteins was not significantly altered in this model. PMCA4 protein could not be detected in isolated VSMCs (A.) and aorta (B.) from PMCA4KO mice by immunohistochemistry (representative images from 3 experiments). PMCA4 protein was principally detected in VSMCs not the endothelium of aorta from PMCA4WT mice (C.), (representative images from 3 experiments). Nuclei stained with DAPI shown as blue in images. Western blot analysis showed aortic protein expression of PMCA1 and NCX1 was similar in PMCA4WT and PMCA4KO mice (D. Mean ± SEM, n=6 & 6 and 5 & 5 respectively). WT KO
PMCA1
Tubulin
100
250
150 50 WT KO
150
NCX1
100 50
Relative expression
(arbitrary units)
150
100
Tubulin
PMCA4WT
PMCA4KO 50
PMCA4WT
PMCA4KO