Atg7 expression is correlated with associated chemoresistance in AML cells. (A) ShControl, stable Atg7 knockdown, and stable Atg7 knockdown transiently overexpressing Atg7 cells were treated with Ara-C (2 μM) and Ida (5 ng/mL) for 72 hours. The flow cytometry was used to measure apoptosis. Whole cell lysate was subjected to immunoblotting for Atg7 (box). β-Actin was used as loading control. (B) Whole cell lysates of AML cell lines (THP1, OCI-AML2 and 3, HL-60, MOLT-4, MOLM 13 and 16) separated on SDS gel electrophoresis and immunoblotted with Atg7 (left panel). HL60 and OCI-AML3 cells were treated with Ara-C (2 μM) and Ida (5 ng/mL) for indicated time periods. The whole cell lysates were immunoblotted with cleaved PARP and cleaved caspase-3 (right panel). Tubulin was used as loading control. Apoptosis was measured using flow cytometry, and autophagy was measured by CytoID kit (bottom table). (C) Ectopic Atg7-expressed HL60 cells were treated with Ara-C (2 μM) and Ida (5 ng/mL) for 48 hours. Lysates were immunoblotted with Atg7, cleaved caspase-3, and cleaved PARP-specific antibody (left panel). β-Actin was used as the loading control. The apoptosis (right panel) was measured using flow cytometry. The statistical significance between the 2 compare groups was calculated by a standard Student t test.