Figure 3. Genetic knockdown of FASN sensitizes cells to death ligands.
A) HeLa and DU145 cells were infected with shRNA targeting SREBP1 or control sequences and stable populations selected as described in materials and methods Total cellular proteins were isolated and analyzed by SDS-PAGE immunoblotting using antibodies against SREBP1, fatty acid synthase (FASN), 3-hydroxy-3-methylglutaryl-Coenzyme A reductase (HMGCR) and tubulin.
B) HeLa and DU145 cells were infected with shRNA targeting FASN or control sequences and stable populations selected. Total cellular proteins were isolated and analyzed by SDS-PAGE immunoblotting using antibodies against FASN and actin.
C) HeLa and DU145 cells infected with FASN or control shRNA as in (B) were treated with CH-11 (100 ng/mL for 18 hours), TRAIL (100ng/mL for 3 hours), or buffer control. After incubation, cell growth and viability was measured by the MTS assay.