Figure 3.

nSMase2 upregulation is independent of known transcriptional regulators of nSMase2. (a) MCF7 cells were seeded in 60 mm dishes and transfected with siRNA to AllStars Negative Control (AS), Sp1, Sp3 or both together. After 24 h, cells were treated with vehicle, and 0.2 or 0.6 μM doxorubicin. After 24 h, cells were collected and immunoblotted for nSMase2, Sp1, Sp3 and actin. (b) MCF7 cells were seeded in 60 mm dishes. One hour before stimulation with doxorubicin or vehicle, they were pretreated with N-acetylcysteine (NAC). Cells were collected and immunoblotted for nSMase2 and actin. (c) MCF7 cells were seeded in 60 mm dishes and transfected with siRNA to AS or CREB3L1. After 24 h, they were treated with vehicle or 0.6 μM doxorubicin. Cells were collected and immunoblotted for nSMase2 and actin