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The data demonstrate the activation by deoxygenation of nonspecific cation channel activity in the SAD mouse red cell membrane.
SAD mouse red cell membrane patches sustained GΩ seals during transitions from room air to nitrogen, and then back to room air.
As shown in Figure 2B, deoxygenation increased mean NPo in SAD mouse red cell patches from 0.01±0.013 to 0.48±0.20 (n = 6; p<0.05).
Buffy coat-depleted cells were washed 5 times in standard mouse red cell wash solution containing (in mM) 172 choline Cl, 10 sucrose, 10 Na Tris-MOPS, pH 7.4, resuspended in storage solution, and kept at 4°C until use.
The on-cell patch records of Figure 1A, with NaCl in both pipette and bath, show that deoxygenation activates noisy channel activity in the SAD sickle mouse red cell membrane.
None of 4 cases of prolymphocytic leukaemia showed mouse red cell rosetting.
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This was different from RBUP which agglutinated rabbit red cells instead of mouse red cells.
NEM-stimulated Cl−-independent K+ efflux of mouse red cells is insensitive to ouabain and bumetanide, but partially inhibited by chloroquine, barium, and amiloride.
Deoxygenation-induced currents in SAD mouse red cells were completely reversible.
Human AA red cells and normal mouse red cells exhibited no deoxygenation-induced increases in current or in [Ca2+]i.
The absence of KCa3.1 in otherwise normal mouse red cells had no effect on the lack of deoxygenation-sensitive [Ca2+]i elevation.
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