Supplementary Figure 6 : K209 is responsible for PEX5 interaction with p62 in response to ROS.

From: ATM functions at the peroxisome to induce pexophagy in response to ROS

Supplementary Figure 6

(a) Representative image of HepG2 cells treated with or without H2O2 for 3 h and immunostained for FIP200 or ATG16L (green) and peroxisomal marker PMP70 (red). Scale bar, 10 μm. High-magnification images of boxed areas showed co-localization between FIP200 or ATG16L and PMP70 (Scale bar, 3 μm). (b) HEK293 cells were transfected with Myc-PEX5-WT and HA-p62, and treated with 0.4 mM of H2O2 for indicated times. The complex of HA-p62 and Myc-PEX5 was immunoprecipitated with anti-HA and immunoblotted with anti-Myc. Inputs were immunoblotted using the indicated antibodies. (c) HEK293 cells were transfected with HA-p62 and Myc-PEX5-WT or Myc-PEX5-K209R. The complex of HA-p62 and Myc-PEX5 was immunoprecipitated with anti-HA and immunoblotted with anti-Myc antibodies. Input lysates were immunoblotted using the indicated antibodies. Uncropped images of western blots are shown in Supplementary Fig. 9.