Fig. 2. Unbiased identification of the target protein of methotrexate (MTX). (a) Representative images from TS-FITGE experiment. Images of Cy3 channel (green, proteome treated with DMSO) and Cy5 channel (red, proteome treated with MTX) were overlaid. (b) Box plot showing the distribution of Cy5/Cy3 fluorescence ratio for each spot in the 53 °C gel. The whiskers indicate 1–99 percentiles. The spot denoted by a triangle in (a) is indicated by a red arrow. (c) Melting curves showing a denaturation pattern of the indicated spot in (b). (d) Cellular thermal shift assay with dihydrofolate reductase (DHFR) antibody.
