Figure 6.

Type of cell death induced by CLytA-DAAO. (A) SW-620, IMIM-PC-2, HGUE-GB-37, and HGUE-GB-39 cell lines were treated with 2 U mL⁻¹ free CLytA-DAAO, and 1 mM D-Ala in the presence or absence of 25 µM Z-VAD for 24 h and cell cycle distribution was determined by flow cytometry. (B) HGUE-GB-18, HGUE-GB-37, and HGUE-GB-39 glioblastoma cell lines were treated with 2 U mL⁻¹ CLytA-DAAO, and 1 mM D-Ala, for 24 h and apoptotic cell death was determined through Annexin V-PE (+) and 7-AAD (−) labeling by flow cytometry. (C) IMIM-PC-2 cell line was treated with 2 U mL⁻¹ CLytA-DAAO, and 1 mM D-Ala in the presence or absence of 10 µM chloroquine (CQ), 10 µM spautine-1 (SP), 20 µM necrostatine-1 (NC), and 10 µM ferrostatin-1 for 24 h and cell cycle distribution was determined by flow cytometry. Data represent the percentage of cells in sub-G1 phase, normalizing the treatment with CLytA-DAAO as 100% ± SEM with n ≥ 3. ** indicates p < 0.01.