FIG 6.

HPV-16 E6 and E7 RGNs rescue p21 and Rb expression in SiHa cells. SiHa cells were transfected with an S. pyogenes Cas9 expression vector and the HPV-16-specific sgRNA constructs indicated and processed for Western blotting. These data are representative of data from 3 biological replicates. (A) Lysate probed for the Flag-tagged Cas9 protein, the p53 effector protein p21, and endogenous β-actin (B). Similar to panel A but with the lysate being probed for Rb expression. The N.S. (nonspecific) sgRNA used as a control is E6 sgRNA1, which is specific for the HPV-18 E6 gene but is not predicted to recognize the HPV-16 E6 gene.