Journal: Virology
Article Title: The HPV16 E7 oncoprotein increases the expression of Oct3/4 and stemness-related genes and augments cell self-renewal.
doi: 10.1016/j.virol.2016.09.020
Figure Lengend Snippet: Fig. 5. Expression of the stem cell-related genes Sox2, Nanog, and Fgf4 in cells expressing the HPV16 E7 oncoprotein. (A and B) A real-time RT-PCR (RT-qPCR) analysis was performed to determine the mRNA expression levels of Sox2, Nanog and Fgf4 in PHK and Saos-2 cells. The RT-qPCR experiments were performed with three in- dependent cultures, and the values are expressed as the means 7 SD (normalized to β2M). (C and D) Western blot analysis of Sox2, Nanog and Fgf4 proteins in PHK and Saos-2 cells. Western blot experiments were performed with three independent cultures, and the expression levels were normalized to those of β-actin. A representative experiment is shown. (E) E7 mRNA expression in PHK and Saos-2 cells stably transfected with E7. The black arrows indicate the expression of E7 and β2M mRNAs in monolayer cell cultures. The analysis was performed in cells that were stably transfected with the HPV16 E7 oncogene. A t test was used to assess the differences in the protein expression levels. Statistical significance was determined as Po0.05. PHK: primary human keratinocytes.
Article Snippet: The Oct3/4 antibody (ab19857, Abcam, USA) was used at a dilution of 1:500, and Sox2 (Sc-20088, Santa Cruz Biotechnology, USA), Nanog (Sc-134218, Santa Cruz Biotechnology, USA), Fgf4 (Sc-1361, Santa Cruz Biotechnology, USA) and β-actin (Sc-130617, Santa Cruz Biotechnology, USA) antibodies were used at a dilution of 1:1,000.
Techniques: Expressing, Quantitative RT-PCR, Western Blot, Stable Transfection, Transfection