Figure 1

Knockdown of Bcl-3 in HeLa cervical cancer cells. HeLa cells were infected with retroviral particles packaged with constructs derived from pSIREN vector (Clontech). These vectors codifies for a double-stranded short hairpin RNA (shRNA) directed toward Bcl-3 under the human U6 promoter. Cells were selected with puromycin for 1 week and protein (upper panel A) or RNA (lower panel A) analyzed. B) HeLa cells were grown in DMEM plus 8% fetal bovine serum for the times shown and viability assessed with the crystal violet method. Mean and standard deviation of the optical absorbance are shown. Black bars: cells expressing a control (scrambled) shRNA, white bars: cells expressing a Bcl-3 shRNA. C) Clonogenic assays. 2.5 × 10³ cells were seeded in 100 mm petri dishes, cultivated for 16 days, fixed and stained with crystal violet. Colonies were manually counted. D) HeLa cells were grown in DMEM plus 8% fetal bovine serum, exposed to ultraviolet B light (280 nm) for 45 seconds (≈ 300 J/m² ) and viability assessed by crystal violet staining 24 hours later. Mean and standard deviation of the optical absorbance are shown. Black bars: cells expressing a control (scrambled) shRNA, white bars: cells expressing a Bcl-3 shRNA. All the assays were performed by triplicate in three separated experiments. Ctrl: control. IB: Immunoblot. *p < 0.05, **p < 0.01 using student's T test.