Detection of Z-VAD-FMK binding in CPC-deficient mutants by western blot. (a–c) Parasites' extracts were analyzed for Z-VAD-FMK binding using avidin-HRP. All parasites were exposed (+) or not (−) to HS (a), H2O2 (b) or miltefosine (c), and incubated with biotin-VAD-FMK (+ b-VAD). Protein extracts were derived from WT, cpc KO and cpc KO. (d–f), Total protein extracts from WT, cpc KO and cpc KO/+ cell lines exposed (+) or not (−) to HS (d), H2O2 (e) or miltefosine (f) and incubated with biotin-VAD-FMK (+ b-VAD) were further analyzed for CPC expression using a CPC anti-serum. Black arrows indicate the two major forms of the CPC protein (34 and 38 kDa, respectively)