Fig. 6
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N-acetyl cysteine treatment to PEX1 and PEX13 depleted cells decreases GFP-MAP1LC3B puncta and increases peroxisome abundance but does not change ubiquitination of PEX5. (A) Immunofluorescent images of HenLe cells stably expressing GFP-MAP1LC3B treated with the indicated siRNA and NAC (10 μM) where indicated. Cells were fixed and immunostained for ABCD3 (red). (B) Quantification of GFP-MAP1LC3B density of (A). (n = 3; 30 cells/trial). Error bars represent standard deviation. (C) Quantification of ABCD3 density of (A). (n = 3; 30 cells/trial). Error bars represent standard deviation. (D) Live cell images of HeLa cells treated with the indicated siRNA and ROS probe OxyBURST. (E) Quantification of mean fluorescent intensity of images in (D) (n = 3; 30 cells/trial). Error bars represent standard deviation. (F) IP analysis of HEK293 cells stably expressing HA-ubiquitin. Cells were treated with the indicated siRNA and NAC where indicated. Asterisks denote p-values; *p < 0.05, **p < 0.01. Scale bars: 25 μm (A), 50 μm (D). |