The SUMOylation Pathway Restricts Gene Transduction by Adeno-Associated Viruses
Fig 2
Knockdown of either the E1 or E2 enzymes of the SUMOylation pathway results in an increased infection efficiency of scAAV2-eGFP.
HeLa cells were transfected with four different siRNAs targeting expression of Ubc9 and Sae2, respectively. Forty-eighth later, the cells were infected with scAAV2-eGFP at an MOIGC of 103. Twenty-four h after infection, half of the cells were proceeded for FACS-analysis, the other half analyzed by western blotting for protein levels after siRNA transfections. (a) Relative MFIs of three independent experiments are shown. The MFI was normalized by setting siRNA transfections with ‘AllStars negative control siRNA’ (Qiagen, indicated as ‘scrambled; scr.’) to 100. (b) The percentage of GFP-positive cells representing infected cells of three independent experiments are shown. (c,d) Western blot analysis showing a reduced steady-state level of Sae2 and Ubc9, respectively, after transfection of the four different siRNAs in comparison to cellular actin. The blot shows whole-cell extracts of two independent transfections for each siRNA. (e) Fluorescence of HeLa cells transduced with scAAV2-GFP vectors. Knockdown of Sae2 or Ubc9 was performed 48 h before transduction, respectively.