Fig. 9
From: Reactive oxygen species enhance rAAV transduction by promoting its escape from late endosomes
Role of ROS in rAAV2 transduction by regulating cathepsins B and L. Hela cells were transfected with 10 nM CTSB siRNA, CTSL siRNA, or scrambled siRNA for 24 h, and then pretreated with H2O2 for 4 h. The pretreated Hela cells were subject to transfection using rAAV2-GFP at 1000 vg/cell. A The representative images of Hela cells transfected with CTSL siRNA, CTSB siRNA and pretreated with H2O2 and then transduced with rAAV2-GFP for 48 h. B Quantification of rAAV2 transduction efficiency after CTSB siRNA, CTSL siRNA and H2O2 pretreatment. C After the treatment, the CTSB level was determined using WB assay. D After the treatment, CTSL expression was measured through WB assay. E Quantitative analysis of CTSB expression. F Quantitative analysis of CTSL expression. G Quantification of rAAV2 in late endosomes when pretreated Hela cells were transduced with rAAV2-GFP for 7 h. Data are represented by means ± SD (n = 3). ns p > 0.05; ****p < 0.0001