FIG 12.

Knockdown of DNA repair DNA polymerase η or κ decreases HBoV1 DNA replication significantly. HEK293 cells were transduced with Pol η- or Pol κ-specific shRNA, as indicated, prior to pIHBoV1 transfection. (A and B) Southern blot analysis. At 48 h posttransfection, cells were collected for Hirt DNA extraction. (A) Hirt DNA samples were digested with DpnI and were subjected to Southern blotting using a probe spanning the HBoV1 NS and Cap genes. A representative blot is shown. (B) The levels of mRF DNA in the blot were quantified and normalized to the level of the control group without shRNA application. Means and standard deviations calculated from three independent experiments are shown. P values were calculated using Student's t test (*, P < 0.05; **, P < 0.01; N.S., no statistically significant difference [P > 0.1]). (C and D) Western blot analysis. At 48 h posttransfection, cells were collected and the cell lysates were analyzed for the expression of Pol η (C) and Pol κ (D). β-Actin was also probed as a loading control.