HPV16 oncoproteins inhibit pro-IL-1β levels.

<p>(A) Human keratinocytes transduced with pLXSN or 16E6E7 were stimulated with AIM2 and (B) NLPR3 ligands and both pro-IL-1β and IL-1β from cell lysates or supernatants were analysed by immunoblotting. β-actin was used as a loading control. Densitometry analysis was performed n = 3. (C) Immunoblotting of pro-IL-1β in human keratinocytes transduced with pLXSN or 16E6E7. (D) Cervical cancer cell lines were lysed and immunoblotting for pro-IL-1β was performed. n = 4 (E) Human keratinocytes transduced with pLXSN or 16E6E7 were treated for 24 h with N-CBZ-Leu-Leu-Leu-al. Cells were harvested and p53, E6 as well as pro-IL-1β levels were determined by immunoblotting. Right, p53 densitometry levels were normalized to β-actin. Below, immunoblot analysis of the 16E6 protein. n = 3. (F) RNA was extracted from Human keratinocytes transduced with pLXSN or 16E6E7 and IL-1β transcripts relative expression was determined by RT-qPCR. n = 5. (G) RNA was extracted from patient derived cervical cancer cell lines and IL-1β transcripts were determined by RT-qPCR. n = 6. Panels A-E. Data are representative of n independent experiments performed in triplicate. Shown are the mean ± SEM with ***, P < 0.0001, based on a two way ANOVA test. Panel F P < 0.0001, based on a one way ANOVA test. Panel G Student unpaired T test was performed comparing C33A to CaSki or SiHa. For immunoblotting data, 1 out of 3 experiments is shown.