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. 2018 Jun 20;3(3):e00204-18. doi: 10.1128/mSphere.00204-18

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Copyright © 2018 Smith et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 4 — The small TbTims are associated with TbTim17. (A to D) Two-dimensional blue native/SDS-PAGE of digitonin-solubilized mitochondrial extracts from (A) wild-type, (B) TbTim9-Myc, (C) TbTim10-Myc, and (D) TbTim8/13-Myc cells followed by immunoblot analysis performed with anti-myc, anti-TbTim17, and anti-CytC1 antibodies. Respective molecular weight markers for the blue native gels (1D-BN-PAGE) and SDS-PAGE gels (2D–SDS-PAGE) are shown. (E to H) Coimmunoprecipitation of digitonin-solubilized mitochondrial extracts from (E) wild-type, (F) TbTim9-Myc, (G) TbTim10-Myc, and (H) TbTim8/13-Myc cells using agarose beads coupled with the anti-myc antibody. Bound proteins were analyzed on SDS-PAGE followed by immunoblot analysis using antibodies for the myc epitope, TbTim17, TbAAC, VDAC, mtHsp70, and TAO. Input (In) lanes were loaded with 10% (vol/vol) of total solubilized proteins; precipitate/bound (Ppt) lanes were loaded with 50% (vol/vol) of the total pulldown fraction. Experiments were repeated three times, and representative results from one trial are shown.