Research Paper Volume 11, Issue 1 pp 33—47

HPV-18 E2 protein downregulates antisense noncoding mitochondrial RNA-2, delaying replicative senescence of human keratinocytes

Figure 1. E2-expressing HFK downregulate the ASncmtRNAs. HFK were transduced in triplicate for 72 h with HPV-18 E2 (HFK-E2) or with control lentivirus (HFK-ZsG) or mock-transduced. (A) Representative analysis of HFK-ZsG and HFK-E2 populations displayed 92% and 78% transduction, respectively. (B) Only HFK-E2 cells expressed the full-length E2 mRNA (1,100 bp). (C) Only HPV-18 E2-transduced HFK expressed E2 protein as evaluated by immunofluorescence (Bars = 40 µm). (D) Relative levels of ncmtRNAs were determined at p3 by RT-PCR using 18S rRNA as loading control. Numbers on the right denote amplicon size in bp. Triplicate analysis of ASncmtRNA-1 (E), ASncmtRNA-2 (F) and SncmtRNA (G) showed that both ASncmtRNAs were downregulated by E2 expression (*p<0.01), while SncmtRNA levels remained unchanged.