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. 2017 Nov 8;7(11):170111. doi: 10.1098/rsob.170111

Figure 2.

Figure 2.

PIR transcripts and protein are overexpressed after ectopic expression of HPV-16 E6/E7 oncogenes in oral epithelial cells and in HR-HPV-positive cervix–uterine cancer cells. (a) PIR mRNA in OKF6-Tert 2 cells stably transfected with and empty vector and pLXSNHPV16E6E7 vector. (b)(i) Western blot for pirin detection in stably transfected OKF6–Tert2 cells; (ii) analysis by densitometry. (c) IF by confocal microscopy for pirin in OKF6–Tert2 cells stably transfected with pLXSNHPV16E6E7 or empty vector and Ca Ski cells. (d) C4I cervical cells were transiently transfected with different concentrations of pLXSNHPV16E6E7 vector, harvested after 24 h (80% confluence) and PIR transcripts were measured by RT–qPCR. (e) Cervical cancer cells (C33A, SiHa and Ca Ski) that contains 0, 2 and 500 copies of HPV-16 were analysed for PIR transcripts expression using RT–qPCR. (f) (i) Western blot for C33A, SiHa and Ca Ski protein extracts using antibodies for pirin and B-actin; (ii) densitometric analysis of western blot bands. Experiments were repeated at least three times with similar results. These results are representative of three independent experiments. **p < 0.01; ***p < 0.001.