Fig. 3. HpPex20p is not required for oligomerisation of AMO and thiolase. (A) Western blot analysis of crude cell extracts revealed that the levels AMO and thiolase were similar in WT and Hppex20 cells. Equal amounts of protein were loaded per lane. (B-F) The oligomeric state of AMO (B), thiolase (C), AO (D), catalase (CAT; E) and HpPex20p (F) were determined by gel filtration analysis. Western blotting of FPLC elution fractions of WT and pex20 cell free extracts revealed similar native sizes for AMO (150 kDa; B), thiolase (100 kDa; C), AO (600 kDa; D) and catalase (240 kDa). In crude extracts of WT H. polymorpha cells HpPex20p was estimated to be 180 kDa (F). WT and Hppex20 cells were grown to the late exponential growth phase on glucose/methylamine for the detection of AMO, CAT, AO and HpPex20p (Fig. 3A,B,D-F) or shortly induced on oleic acid/ammonium sulphate to induce thiolase (A,C).