Fig. 5.

PCI-24781 displays a direct apoptotic effect in GBM cells. A U-118MG cells were treated with 1.25 μM PCI-24781 for 24 and 48 h, and cell lysates were analyzed for pro and anti-apoptotic markers by western blotting. B-D PCI-24781 induces ROS generation in GBM cells. B U-118MG cells were treated with vehicle control, TMZ, PCI-24781 or TMZ, and PCI-24781 combination for the indicated times. After incubation, media were removed, and cells rinsed with Krebs-Ringer buffer and stained with 10 μM DCF-DA at 37^•C for 30 min in the dark. Finally, cells were washed in buffer, and fluorescent images were taken, and the representative images are shown. Scale bar 400 μm. C U-118MG cells were incubated with drugs for 6 h, and lysates were analyzed for lipid peroxidation marker 4-HNE by immunoblotting. D U-118MG cells were incubated with vehicle control, PCI-24781, PCI-24781 + NAC (15 mmol/L), or 2 h pre-treatment with NAC, followed by PCI-24781 for 48 h, and lysates were analyzed for apoptotic markers. Acetylation of α-tubulin serves as the marker for HDAC inhibitor PCI-24781 treatment. ROS- reactive oxygen species; NAC- N-acetylcysteine