Figure 5.

Subcellular fractionation shows that Par3 is redistributed in HPV‐positive cells in an E6‐dependent manner. HeLa cells were transfected with siRNA against either luciferase or E6.18. After 48 hs, cells were harvested and protein extracts were fractionated into cytosolic, membrane, nuclear and cytoskeletal fractions. The expression of Par3 and markers for the four subcellular fractions were assessed by western blotting. Expression of vimentin, p84, α‐tubulin, and transferrin receptor (TR) were used as control for the integrity of the cytoskeleton, nucleus, cytosolic and membrane fractions, respectively. The ablation of E6.18 was monitored by the expression of p53 in the nuclear fraction. The results shown are representative of three independent experiments. Numbers are folds of band intensity for Par3 in E6 depleted cells with respect to the luciferase siRNA control (considered as 1). The intensity of each band was normalized to the corresponding loading control in the different subcellular fraction.