Journal: Nature cell biology

Article Title: ATM Functions at the Peroxisome to Induce Pexophagy in Response to ROS

doi: 10.1038/ncb3230

Figure Lengend Snippet: ( a ) HEK293 cells expressing Myc-PEX5-WT and HA-Ub treated with H 2 O 2 (0.4 mM) for 6 h, were co-immunoprecipitated using anti-HA and blotted using anti-Myc antibody. The inputs were immunoblotted using the indicated antibodies. ( b ) HEK293 cells expressing Myc-PEX5-WT and HAUb-WT or HA-Ub-K0 (ubiquitin with all lysine residues mutated to arginine) constructs were treated with H 2 O 2 (0.4 mM) for 6 h, and Myc-PEX5 co-immunoprecipitated using anti-HA and blotted using anti-Myc antibody. Inputs were immunoblotted using the indicated antibodies. ( c ) Schematic representation of the N-terminus of PEX5 showing the putative ATM phosphorylation site (S141) and ubiquitination site (K209). ( d ) HEK293 cells expressing HA-Ub-WT and Myc-PEX5-WT or Myc-PEX5-K209R plasmids were treated with H 2 O 2 (0.4 mM) for 6 h, and Myc-PEX5 coimmunoprecipitated using anti-HA antibody, and blotted using anti-Myc antibody. Input lysates were immunoblotted using the indicated antibodies. ( e ) HEK293 cells expressing HA-Ub-WT and Myc-PEX5-WT or Myc-PEX5-S141A constructs were treated with H 2 O 2 (0.4 mM) for 6 h, and Myc-PEX5 co-immunoprecipitated using anti-HA antibody and blotted using anti-Myc antibody. Input lysates were immunoblotted using the indicated antibodies. Uncropped images of western blots are shown in .

Article Snippet: HA-Ub (#18712) and HA-Ub-K0 (#17603) plasmids were purchased from Addgene.

Techniques: Expressing, Immunoprecipitation, Construct, Western Blot