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. 2019 Apr 25;14(5):1020–1029. doi: 10.1021/acschembio.9b00222

Figure 2.

Figure 2

Identification of ClpP as an ONC201 and TR compound binding protein. (a) HeLa cell lysate was mixed with the indicated beads in the absence or presence of free TR-80. A silver stained gel is shown, and the band indicated by the arrow was excised and subjected to mass spectrometry for identification. (b) Replicate samples from panel a were immunoblotted with ClpP antibodies. (c, d) TR-81 beads were charged with HeLa cell lysate and eluted with 50 μM TR-57. A portion of the eluant was silver stained (c) or immunoblotted (d) for ClpP. (e) Pure, recombinant human ClpP was mixed with the indicated beads in the absence or presence of 50 μM TR-57 and immunoblotted for ClpP. (f, g) HeLa cell lysates were mixed with TR-81 beads in the presence of the indicated compounds and immunoblotted for ClpP. (h) HeLa cells were treated with the indicated compounds for 30 min, and lysates were mixed with TR-81 beads and immunoblotted for ClpP.