Suppression of eIF4AI/II in HeLa cells leads to inhibition of proliferation and increased cell death. (A) Western blot analysis of eIF4AI/II in HeLa cells upon siRNA transfection. Antibodies and siRNAs used are indicated to the right and top of the panels, respectively. siNT indicates nontargeting siRNA. α-eIF4AI, sc-14211. (B) Western blot analysis of eIF4AI/II in HEK293, A549 (lung adenocarcinoma), KMS-11 (multiple myeloma), and SK-N-BE (neuroblastoma) cells transfected with the indicated siRNAs. α-eIF4AI, ab31217. (C) Consequences of eIF4AI/II suppression on HeLa cell proliferation. Cells were transfected with the indicated siRNAs, plated at a density of 12,000 cells per well in a 12-well plate, and cell numbers were monitored by counting. Each value represents the average of triplicate samples from three independent experiments. Error bars, SEM. (D) Cell cycle analysis of HeLa cells transfected with the indicated shRNAs. Cell cycle analysis was performed 72 h following transfection with the indicated siRNAs. Error bars, SEM. n = 3. (E) EIF4AI suppression increases cell death. HeLa cells were harvested 48 h after siRNA transfection, and cell death was quantitated as the percentage of PI+ cells measured by flow cytometry. n = 6; (*) P < 0.0001.
