Fig. 3.

BAG3^P209L forms soluble oligomers. a Replacing proline for leucine at residue 209 is predicted to increase local beta-sheet content. PepFold was used to estimate propensity to form secondary structure. b Solutions containing recombinant BAG3^wt or BAG3^P209L were stained with SYPRO orange and fluorescence was measured. Results are the average of at least three independent experiments performed in triplicate each. Error bars represent SD. c Recombinant BAG3^wt or BAG3^P209L solutions were labeled by the beta-sheet dye, thioflavin T. Results are the average of at least three independent experiments performed in triplicate each. Error bars represent SD. d BAG3^P209L does not have gross changes in secondary structure, as estimated from CD measurements. Results are representative of measurements performed in triplicate. e BAG3^wt or BAG3^P209L solutions were subjected to partial proteolysis with either chymotrypsin or trypsin and samples were loaded on SDS–PAGE for separation. Coomassie stained gels are shown. f Fractionation of HEK293 cells expressing FLAG-BAG3^wt or FLAG-BAG3^P209L over a sucrose gradient. Western blot against BAG3 and GAPDH is shown. Source data are provided as a Source data file