Journal: bioRxiv

Article Title: Pharmacological activation of PIEZO1 in human red blood cells prevents Plasmodium falciparum invasion

doi: 10.1101/2021.05.28.446171

Figure Lengend Snippet: Haemolytic activities of the PIEZO1 activators Yoda1, Jedi1 and Jedi2. Blood of different donors was cultured for 24h at 2.5% haematocrit in standard culture conditions (RPMI without phenol red with 0.5% albumax) in the presence of the indicated concentrations of compounds. Haemoglobin released into the culture supernatant was quantified by measuring the absorbance at 540 nm in a TECAN spectrophotometer and expressed as percentage of total lysis induced by the addition of 0.15% saponin. Shown are mean and SD, n = 5 for Yoda1 and n = 4 for Jedi1 and Jedi2, all performed in triplicates.

Article Snippet: The following chemicals and their stock concentrations in DMSO (in square brackets) were used in this study: Yoda1 (Sigma-Aldrich)[20 mM], A23187 (Sigma-Aldrich)[2 mM], Jedi1 (MolPort, Riga, Latvia)[200 mM], Jedi2 (MolPort)[200 mM], Nystatin (Sigma-Aldrich)[40 mg/ml].

Techniques: Cell Culture, Spectrophotometry, Lysis

Journal: bioRxiv

Article Title: Pharmacological activation of PIEZO1 in human red blood cells prevents Plasmodium falciparum invasion

doi: 10.1101/2021.05.28.446171

Figure Lengend Snippet: PIEZO1 activation in RBCs by Jedi1 and Jedi2. ( A ) The effect on parasitemia was observed either in the continuous presence of drug (drug on) in complete medium, or after a pre-treatment for 10 min in RPMI followed by extensive washes (drug off) before adding synchronized schizont infected RBCs for culture in complete medium. Parasitemia was determined after incubation for 18 h and expressed as percentage of mock-treated controls (n=3, mean + SD). Kruskal-Wallis test for drug on condition of Jedi1 and Jedi2 p<0.05. ( B ) Cell shape of the cells at 4 min after addition of compounds (5 μM Yoda1, 1 mM Jedi1 and Jedi2). Scale bar = 5 μm. ( C ) Osmotic fragility assays with RBCs treated with increasing concentrations of Jedi1 (top) and Jedi2 (bottom). One representative experiment (of 3) is shown.

Article Snippet: The following chemicals and their stock concentrations in DMSO (in square brackets) were used in this study: Yoda1 (Sigma-Aldrich)[20 mM], A23187 (Sigma-Aldrich)[2 mM], Jedi1 (MolPort, Riga, Latvia)[200 mM], Jedi2 (MolPort)[200 mM], Nystatin (Sigma-Aldrich)[40 mg/ml].

Techniques: Activation Assay, Infection, Incubation