Figure 4.
Constitutively active Akt protects MCA-105 cells from the cytotoxic effect of MJ. (A) MCA-105 cells were seeded into 24-well plates at a concentration of 0.4 x 105 cells per well and were transfected with GFP, myr-Akt, or myr-KD-Akt plasmids. Forty-eight hours after transfection, total cell lysates were subjected to SDS-PAGE followed by immunoblot analysis using anti-Akt antibodies. (B) MCA-105 cells were seeded into 24-well plates at a concentration of 0.4 x 105 cells per well and were transfected with either myr-Akt or myr-KD-Akt plasmids. Forty-eight hours after transfection, vehicle (ethanol) or MJ (3 mM) was added to the wells for 2.5 hours. Cells were collected for FACS analysis to assess cell death with the PI reagent as described in the Materials and Methods section.