FIGURE 1.

Alternol interacts with multiple cellular proteins. A, PC-3 cells were teated with the solvent DMSO, biotin, Alternol, biotin-labeled Alternol (10 μM) for 4h. Cell lysates were incubated with Streptavidin agarose resin overnight at 4°C. After three washes, the elutes were run on SDS-PAGE followed by comassine blue staining. Protein bands were dissected for Mass-Spectrometry. B, The Biotin-Streptavidin pulldown elutes were used for Western blots with antibodies as indicated. C, Both pulldown elutes and whole cell protein lysates were used for the Western blots. Caspase-3 blot was included as a negative target control. D-G, PC-3 cells were treated with the solvent DMSO or Alternol for 4 h and cellular proteins were extracted for CETSA assay as described in the text. Protein band densities were acquired with ImageJ software and the curve plots were generated with Graphpad Prism 5.0 software