Figure 4.
Figure 4. PSAC activity on RBCs from CFTRΔF508/ΔF508 donors. (A) Single-channel recordings on an RBC from a CFTRΔF508/ΔF508 donor infected with a trophozoite-stage parasite. Bath and pipette contained solution A. Vm was +100 or -100 mV (upper and lower traces, respectively). Closed channel levels are indicated by dashed red lines. (B) Recording from a 2-channel patch on an infected CFTRΔF508/ΔF508 RBC in solution A at a Vm of -100 mV. Downward transitions represent the opening of either one or both channels; corresponding levels are marked with dotted lines. (C) Whole-cell recording on an infected CFTRΔF508/ΔF508 RBC with near physiologic bath and pipette solutions of 165 mM NaCl, 10 mM MgCl2, 5 mM CaCl2, 20 mM Na-HEPES, pH 7.4, before or after addition of 200 μM furosemide (left and right groups, respectively). (D) Whole-cell currents with solution A in bath and pipette from infected RBCs using healthy (left traces) and CFTRΔF508/ΔF508 donors (right traces). In panels C and D, traces for 50-ms steps to Vm between -100 mV and +100 mV in 20-mV increments are superimposed to demonstrate PSAC's voltage dependence; the zero current levels are marked with horizontal red lines. Scale bars represent 2.9 pA/127 milliseconds (A), 3.1 pA/500 ms (B), 1590 pA/30 ms (C), and 3000 pA/30 ms (D). Similar PSAC activity was recorded from each of 6 CFTRΔF508/ΔF508 donors after in vitro parasite culture.

PSAC activity on RBCs from CFTRΔF508/ΔF508 donors. (A) Single-channel recordings on an RBC from a CFTRΔF508/ΔF508 donor infected with a trophozoite-stage parasite. Bath and pipette contained solution A. Vm was +100 or -100 mV (upper and lower traces, respectively). Closed channel levels are indicated by dashed red lines. (B) Recording from a 2-channel patch on an infected CFTRΔF508/ΔF508 RBC in solution A at a Vm of -100 mV. Downward transitions represent the opening of either one or both channels; corresponding levels are marked with dotted lines. (C) Whole-cell recording on an infected CFTRΔF508/ΔF508 RBC with near physiologic bath and pipette solutions of 165 mM NaCl, 10 mM MgCl2, 5 mM CaCl2, 20 mM Na-HEPES, pH 7.4, before or after addition of 200 μM furosemide (left and right groups, respectively). (D) Whole-cell currents with solution A in bath and pipette from infected RBCs using healthy (left traces) and CFTRΔF508/ΔF508 donors (right traces). In panels C and D, traces for 50-ms steps to Vm between -100 mV and +100 mV in 20-mV increments are superimposed to demonstrate PSAC's voltage dependence; the zero current levels are marked with horizontal red lines. Scale bars represent 2.9 pA/127 milliseconds (A), 3.1 pA/500 ms (B), 1590 pA/30 ms (C), and 3000 pA/30 ms (D). Similar PSAC activity was recorded from each of 6 CFTRΔF508/ΔF508 donors after in vitro parasite culture.

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