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Blood, Vol. 110, Issue 6, 2158-2165, September 15, 2007
Point mutations involved in red cell stomatocytosis convert the electroneutral anion exchanger 1 to a nonselective cation conductance
Blood Guizouarn et al.
110: 2158
Supplemental materials for Guizouarn et al, Vol. 110, Issue 6, 2158-2165
Files in this Data Supplement:
- Figure S1. Na+ and K+ contents in oocytes expressing the different hAE1 point mutations in the presence or absence of ouabain and bumetanide in incubation media (PDF, 12.6 KB) -
Na+ (white bars) and K+ (black bars) were measured in oocytes five days after injection in (A) MBS or in (B) MBS with 0.5 mM ouabain and 5 µM bumetanide. Data presented are means plus and minus the SEM of 15 oocytes from the same batch and are representative of 3 experiments.
- Figure S2. Comparison of I/V curves obtained with oocytes expressing wt hAE1 and mutant hAE1 in different media (PDF, 34.1 KB) -
(A) Current-voltage curves were measured in MBS where Na+ was totally replaced by the impermeant cation NMDG. The basal conductance of oocytes injected with wild type hAE1 is plotted by the dotted line, the conductance of oocytes expressing the different point mutations are in the plain line. (B) Current-voltage curves measured in MBS where Na+ was substituted by K+. Dotted lines represent the conductance of control oocytes expressing wt hAE1, and plain lines represent the conductance of oocytes expressing the different point mutations of hAE1. Data are means of n oocytes coming from different batches (in NMDG medium, n=10 for L687P, 11 for D705Y, 15 for S731P, 13 for H734R and 14 for wt. In KCl medium, n=10 for L687P, D705Y, S731P, 11 for wthAE1 and 16 for H734R).
- Figure S3 Analyses of currents induced by co-expression of hGPA and S731P mutation of hAE1 (PDF, 15.8 KB) -
(A) SITS sensitivity of currents recorded in oocyte co-expressing the S731P mutation with hGPA. Black diamonds refer to oocytes incubated in MBS and white diamonds refer to oocytes incubated in MBS with 10−4 M SITS. The same oocytes were used to record I/V curves in absence or presence of SITS. This figure shows one oocyte representative of 10. (B) A batch of five oocytes co-injected with hGPA and S731P mutation was used 3 days after injection to record I/V curves in MBS (black diamonds), in NMDG medium (NaCl substituted by NMDGCl, white diamonds), or in KCl medium (NaCl substituted by KCl, grey triangles). Data are means plus and minus a SEM. of 5 oocytes. When not seen, SEM was smaller than the symbol.
- Figure S4. Effects of Zn2+ or La3+ on currents induced by co-expression of H734R and hGPA (PDF, 14.2 KB) -
Black diamonds are the current values in MBS (without bicarbonate and sulfate to avoid precipitation with inhibitors used). White diamonds are the current values in the presence of 1 mM Zn2+ (A) and white triangles are currents in presence of 1 mM La3+ (B). Data are means plus and minus a SEM. of 5 oocytes from two different experiments.
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