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. 2022 Apr 11;13(2):e03849-21. doi: 10.1128/mbio.03849-21

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Copyright © 2022 Santos 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 — Anti-HA bead immunoprecipitation (IP) of mock-HA and HA-ETMP1 strains. (A) Western blot analysis using anti-HA antibody of the cell lysates and various IP fractions of mock-HA (left) and HA-ETMP1 (right). A black arrowhead indicates the position of HA-tagged ETMP1 (33 kDa). (B) Silver-stained-SDS-PAGE gel of IP eluates of mock-HA and HA-ETMP1 strains. A black arrowhead points to a specific ∼55-kDa band unique to HA-ETMP1. (C) Enriched or exclusively detected proteins in the ∼55-kDa excised gel band from HA-ETMP1 IP eluate compared to that of mock-HA control IP eluate by liquid chromatography-tandem mass spectrometry (LC-MS/MS) sequencing analysis. MW, predicted molecular weight; Qv, quantitative value (normalized total spectra). The presence of the detected proteins in the previously published mitosome proteome data (18) was analyzed, and the results are listed in the last column (+, present; −, absent). (D) Total cell lysates of mock-HA and HA-ETMP1 were separated by BN-PAGE, followed by anti-HA Western blot analysis. Black and red arrowheads indicate the ∼180-kDa and ∼90-kDa complexes, respectively, that contain HA-ETMP1.