Figure 7. LSD1 inhibition and autophagy blockade exert synergistic effects on cancer cell apoptosis.

(A) Uterine serous carcinomaARK2 cells were treated with 100nM SP2509 and 25nM chloroquine for 24 h. (B) ARK2 cells were transiently transfected with si-C or LSD1 siRNA (si-LSD1) for 48 h and subsequently treated with 25nM chloroquine for 24 h. Equal amount of protein lysates were subjected to immunoblotting with the indicated antibodies. GAPDH was used to confirm equal protein inputs in all lanes. (C) ARK2 cells were treated with 100nM SP2509 and 25nM chloroquine for 72 h Equal amount of protein lysates were subjected to immunoblotting with the indicated antibodies. β-actin was used to confirm equal protein inputs in all lanes. (D) Cell survival was analyzed with MTT assays. (E) ARK2 cells were subcutaneously injected into the lateral hind leg of nude mice. Xenografted tumors were treated with subcutaneous injections of vehicle (n = 4), SP2509 (n = 4), chloroquine (CQ) (n = 4), or the combination of SP2509 and chloroquine (SP2509+CQ) (n = 4) for 4 weeks. Tumor diameter was measured weekly and tumor volume (cm³) was calculated. * P < 0.05 compared to the control group. (F) Representative tumors were taken from tumor-bearing nude mice treated with vehicle, SP2509, CQ or SP2509 +CQ. (G) Tumors treated with vehicle, SP2509, CQ or SP2509 +CQ were immunoblotted with the indicated antibodies. GAPDH was used to confirm equal protein input in all lanes.