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. Author manuscript; available in PMC: 2007 Feb 16.
Published in final edited form as: Oncogene. 2006 Mar 30;25(14):2094–2104. doi: 10.1038/sj.onc.1209244

Figure 3.

Figure 3

Stable expression of HPV16 E7 siRNA in HPV16+ cervical cancer cells. (a) Diagram of the HPV16 E7 siRNA expression vector, a pSUPER-derived pST66, and the expressed shRNA 198 sequence. BSD is a blasticidin S deaminase gene used as a mammalian selection marker. HPV16+ cervical cancer cells, CaSki and SiHa, transfected with the plasmid pST66 or pSUPER vector control, were grown in the presence of blasticidin S in the culture medium and the blasticidin S-resistant cells were selected to grow into a confluent monolayer for further analysis. Total cell RNA extracted from those cells stably transfected with the plasmid pST66 (CaSki for C-1 to C-3 and SiHa for S-2 to S-3) or with the pSUPER vector only (CaSki for C-V and SiHa for S-V) was tested for the presence of HPV16 E7 siRNA by Northern blot (b). Protein samples extracted from the stable siRNA cells were examined for HPV16 E7 expression by Western blot (c). Sample loading was normalized with U6 snRNA for Northern blot (b) and β-tubulin for Western blot (c).