Fig. 2

Validation of Vimentin as an Ajoene Target. (a) Immunoblot of human recombinant vimentin treated with DP (100 μM) in the absence or presence of DTT (100 mM), probed with a primary anti-dansyl and anti-vimentin (H-84) antibody. (b) Human recombinant vimentin was treated with 100 μM ZA or DP and purified by SDS-PAGE. The band excised from the gel was digested with trypsin and fragments were identified by MS/MS MALDI-TOFF mass spectrometry. The Cys-328 containing fragment qvqsltcevdalk was detected in the control and treated samples carrying a 2+ charge where m/z = [M + 2H]2+. (c) In the samples treated with ZA or DP, the predicted m/z ratio of the modified fragment was observed