Pharmacological inhibition of phosphodiesterases can repair phenotypic defects in the TgATPaseP-GC mutant. Pharmacological inhibition of phosphodiesterases.

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Pharmacological inhibition of phosphodiesterases can repair phenotypic defects in the TgATPaseP-GC mutant. Pharmacological inhibition of phosphodiesterases can repair phenotypic defects in the TgATPaseP-GC mutant. (A) Gliding motility of the Pnative-TgATPaseP-GC-HA3’IT-3′UTRexcised mutant and progenitor (Pnative-TgATPaseP-GC-HA3’IT-3′UTRfloxed) strains in the presence of two known phosphodiesterase inhibitors, zaprinast (500 μM), and BIPPO (55 μM). Fresh syringe-released parasites were treated with drugs for 15 min during the assay, followed by fixation and staining with α-TgSag1 antibody. 500–600 parasites of each strain were evaluated for the motile fraction, and 100–120 trail lengths were measured by ImageJ (n = 3 assays, mean with SEM). (B, C) Egress and invasion rates of indicated parasite strains after exposure to zaprinast and BIPPO. Intracellular and extracellular parasites were differentially stained with α-TgGap45 and α-TgSag1 antibodies, as shown in the Materials and Methods section. For egress, parasitized cells (MOI, 1; 40 h postinfection) were stimulated with either zaprinast (500 μM) or BIPPO (55 μM) for 5 min 30 s before fixation and staining. Drug treatment during invasion assay was performed for 1 h. In total, 500–600 vacuoles and 1,000 parasites were scored for each strain in panels (B) and (C), respectively (n = 3 assays, mean with SEM). Statistics was performed for individual pair of columns using t test (*P ≤ 0.05; **P ≤ 0.01; and ***P ≤ 0.001). Özlem Günay-Esiyok et al. LSA 2019;2:e201900402 © 2019 Günay-Esiyok et al.