Fig. 2.
In vitro pull-down assay. Cell lysates were incubated with purified enhanced green fluorescent protein cocaine- and amphetamine-regulated transcript (EGFPCART) proteins (lanes 1 and 3) or EGFP proteins (2 and 4), pulled down by protein G beads linked to anti-CII (SDHB) antibody, and then probed with either anti-EGFP (A) or anti-CART (B) antibodies (same membrane stripped and reprobed). Lysates in lanes 1 and 2 were prepared from PC3 cells, 3 and 4 from HEK293 cells using CelLytic Reagent. Lane 5 is input loaded with approximately 10% of the EGFPCART protein used in the binding reaction. HC, heavy chain of Ig; LC, light chain of Ig.