Fig. 1.

Anti-AKT and anti-pAKT immunoreactive bands in Leishmania parasites and Leishmania killing by AKT inhibitor X. a L. panamensis (Lp) and L. infantum (Li) promastigotes were cultured in the presence (+FBS) or absence (−FBS) of FBS, and protein extracts were submitted to Western blot using anti-human AKT1/2/3 and anti-pAKT(Thr308) polyclonal antibodies. b L. panamensis (Lp) and L. donovani (Ld) promastigotes were cultured at 26 °C or 37 °C, and protein extracts were submitted to Western blot using anti-human AKT1/2/3 and anti-pAKT(Thr308) polyclonal antibodies. The positions of the immunoreactive bands are indicated by arrows. c Promastigotes of different Leishmania spp. were incubated for 14 h at 26 °C in the presence (+FBS) or absence (−FBS) of FBS, and without (Control) or with 5 μM AKT inhibitor X. Then, parasites were collected and analyzed for the induction of apoptosis-like cell death as assessed by the percentage of parasites in the sub-G₀/G₁ region (hypodiploid cells) by flow cytometry. d Promastigotes of different Leishmania spp. were incubated at 37 °C for 3 h in the presence or absence (Control) of 5 μM AKT inhibitor X. Then, parasites were collected and analyzed for the induction of apoptosis-like cell death as assessed by the percentage of parasites in the sub-G₀/G₁ region (hypodiploid cells) by flow cytometry. e, f J774 macrophage-like cells infected with L. panamensis were untreated (Control) or treated with 10 μM AKT inhibitor X for 24 h. Then, the number of intracellular amastigotes was quantified by Giemsa staining (e), and the percentage of apoptotic macrophages was assessed through the percentage of mammalian cells at the sub-G₀/G₁ region (hypodiploid cells) by flow cytometry (f). Data shown are means ± SD or representative of 3 independent experiments. Asterisks indicate statistically significant differences with respect to control values (**P < 0.01). Western blot experiments shown are representative of 3 performed