Figure 2
Atg7 knockdown enhances chemosensitivity in AML cells. (A) Atg7 knockdown (ShAtg7) or nonsilencing scrambled control (ShControl) OCI-AML3 AML cells were treated with Ara-C (2 μM) or (B) Ida (5 ng/mL) for different time intervals, and total lysates were immunoblotted to detect autophagy. Duplicate samples were stained with autophagy marker LC3-II to quantify autophagy using flow-based assay (A right panel). Similarly, apoptosis was measured using annexin V binding after treatment with (C) Ara-C (2 μM) and (D) Ida (5 ng/mL) for different time intervals, respectively. AML patient samples were either transfected or transduced with siRNA (E) or shRNA (F) targeting Atg7 for 48 hours, and then cells were further treated with Ara-C (2 μM) and Ida (5 ng/mL) for 72 hours. The apoptosis was measured using flow cytometry (annexin binding). Lysates were immunoblotted with Atg7 antibody to confirm knockdown of Atg7 (data not shown). The statistical significance of percent apoptosis in ShAtg7 compared with ShControl was calculated by the Student t test.

Atg7 knockdown enhances chemosensitivity in AML cells. (A) Atg7 knockdown (ShAtg7) or nonsilencing scrambled control (ShControl) OCI-AML3 AML cells were treated with Ara-C (2 μM) or (B) Ida (5 ng/mL) for different time intervals, and total lysates were immunoblotted to detect autophagy. Duplicate samples were stained with autophagy marker LC3-II to quantify autophagy using flow-based assay (A right panel). Similarly, apoptosis was measured using annexin V binding after treatment with (C) Ara-C (2 μM) and (D) Ida (5 ng/mL) for different time intervals, respectively. AML patient samples were either transfected or transduced with siRNA (E) or shRNA (F) targeting Atg7 for 48 hours, and then cells were further treated with Ara-C (2 μM) and Ida (5 ng/mL) for 72 hours. The apoptosis was measured using flow cytometry (annexin binding). Lysates were immunoblotted with Atg7 antibody to confirm knockdown of Atg7 (data not shown). The statistical significance of percent apoptosis in ShAtg7 compared with ShControl was calculated by the Student t test.

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