Figure 2.

Osmotic resistance tests. Fresh venous blood was obtained by venipuncture from informed patients and healthy volunteers in EDTA collecting tubes kept at 4°C until analysis. Experiments were performed 24h after withdrawn. Fresh blood was washed 4 times at room temperature in medium containing in mM: NaCl 147, KCl 5, MgCl₂ 1, CaCl₂ 1, Hepes 10, buffered with NaOH pH7.4 (320 mOsm). RBC suspension (40% hematocrit) was then incubated at 37°C for 18 hours. Blue squares: control at t=18h, red squares: 10 μM TRAM-34 18 h incubation, orange diamonds: 0.4 μM Senicapoc (MedChem Express) 18h incubation, Green triangles: 4 μM Senicapoc 18h incubation, grey circles: 10 μM GsMTx4 (Grammostola spatulata toxin, specific blocker of Piezo1: Bae C. et al. Biochemistry 2011; 50(29)) 18h incubation. The black line with white diamonds represent the osmotic resistance curve done at blood reception, on total blood, before washing in saline buffer and starting the incubation at 37°C. Data are means±sem n=3.