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. 2017 Oct 31;11(12):1788–1805. doi: 10.1002/1878-0261.12145

Figure 3.

Figure 3

Identification of dasatinib‐binding proteins in MCF7 cells. (A) Schematic representation of the chemical proteomics approach used for dasatinib‐binding protein identification in MCF7 cells. (B) Silver‐stained SDS/PAGE of protein–resin complexes. MCF7 cell lysates were incubated with dasatinib coupled to epoxy‐activated Sepharose 6B medium (dasatinib‐resin) or to uncoupled epoxy‐activated Sepharose 6B medium (control resin). Bands of interest were excised from the gel, and the proteins from these bands were analyzed by MALDI‐TOF. Main proteins identified in bands 1–3 across Swiss‐Prot database are shown in the table to the right, together with the proteins' molecular weight and the proteins' score Mascot reported. M r refers to molecular mass (kDa). (C) Western blotting analysis of the interaction of SRC, CSK, YES, FYN, LYN, and LCK with dasatinib by pull‐down experiments with the resins mentioned in (B). Cell extracts were used to detect total protein levels. Data information: In (B,C), results from a representative experiment that was repeated twice are shown.