CYP5122A1, a Novel Cytochrome P450 of Leishmania Figure 8. Effect of drug exposure. A: Bar graph shows percentage viability of WT and HKO parasites in response to treatment with PAT. Mean 6 SE, n = 4; * P#0.05. B: Bar graph shows percentage viability of WT and HKO parasites in response to treatment with miltefosine. Mean 6 SE, n = 4; * P#0.05. C. Western blot analysis (i) and its quantitation (ii) of total cell lysate from Leishmania donovani promastigotes post treatment with different doses of PAT (100 mg/mL, 200 mg/mL and 300 mg/mL) for 24 h probed with anti-CYP5122A1 antibody. D. (i) Western blot analysis of Leishmania total cell lysate post treatment with different concentrations of miltefosine for 24 h, (ii) quantitation of immunoblots in (i); (iii) western blot analysis of L. donovani whole cell lysate upon treatment with 20 mM miltefosine for different time periods. Mean 6 SE, n = 4. E. Levels of intracellular ROS in response to treatment with miltefosine measured using the fluorogenic probe CMH2DCFDA. Mean 6 SE, n = 3. doi:10.1371/journal.pone.0025273.g008 PLoS ONE | www.plosone.org 10 September 2011 | Volume 6 | Issue 9 | e25273