Activation of AKT is perturbed in the absence of CKII phosphorylation of E7.

(A) HEK 293 cells were transfected with HA-AKT together with HPV-16 E7 FLAG-HA, wildtype or 16 E7 S31A/S32A, HPV-18 E7 FLAG, wildtype or 18 E7 S32A/S34A or empty vector. After 24 hours, the cells were serum starved for 16 hours and harvested directly with SDS PAGE sample buffer and analysed by Western blotting for phospho-AKT, total AKT (HA), phoshpo 16E7 and total E7 (FLAG), as indicated. (B) Confluent C4-1 wildtype and CKII mutant cells were serum starved for 48 hours and total levels of AKT, p-AKT, and α-actinin were analysed by Western blotting. (C, D, E) Confluent C4-1, CaSki and C33a cells were either treated with AKT inhibitor or PI3K inhibitor in serum free medium for 48 hours. Conditioned medium from the treatments were analysed by Western blotting for MMP1, MMP13 and MMP8. In each case the lower panels show the Ponceau stain of the nitrocellulose membranes.