Figure 6

Expression silencing of human eIF3f in A549 cells. A) WB analysis of eIF3f expression in transfected pMSG (control) and pFH11A (antisense) A549 cultures. Tubulin WB is shown to document equal protein loading. B) WB analysis of eIF3f expression in transfected scrambbled (control) and eIF3f siRNAs A549 cultures. Tubulin WB is shown to document equal protein loading. C) Difference in cell growth in control (water and scrambled siRNAs) and eIF3f siRNA treated A549 cultures after 24 and 48 hours of transfection. Left: suspended and adherent cells stained with trypan blue to assess cell viability after 24 hours of transfection. Right: Cells were stained with acridine orange and ethidium bromide to visualize cell morphology and death by necrosis or apoptosis. Timepoint cero represents cells before transfection. Final magnification: 40×. D) Percent of apoptosis in control (water and pMSG), antisense eIF3f (pFH11A), scrambled siRNAs and eIF3f siRNAs transfected cells, estimated from cell counts in microscopic fields similar to those shown in panel C. Data expressed as mean ± SE; * p < 0.01 with respect to their respective control (water, scrambled siRNAs or pMSG), n ≥ 3.