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. 2020 Jan 17;9:445. doi: 10.3389/fcimb.2019.00445

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Copyright © 2020 Chasen, Coppens and Etheridge.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Identification of the CP1 associated proteins CP2 and CP3. (A) SDS page gel of CP1-mNeon-Ty CoIP eluate stained for total protein. Several bands are present in the CoIP that are absent from the control IP using parental lysate. Band excision and MALDI mass-spectroscopy (MS) analysis confirmed that the upper band (green arrow) is CP1-mNeon-Ty. (B) Notable hits from Orbitrap shotgun MS of the CP1 CoIP eluate revealed two hypothetical protein (CP2, CP3) in addition to CP1. αTy immunoblots of lysates from CP2-mNeon-Ty (C) and CP3-mNeon-Ty (D) overexpressing epimastigotes show that the tagged protein are expressed. (E,F) SR-SIM of CP2-mNeon-Ty and CP3-mNeon-Ty overexpressing mutants showing SPC labeling. 4°C labeling of with Concanavalin A -Rhodamine labels the SPC pre-oral ridge (white arrows). CP2-mNeon-Ty (G) and CP3-mNeon-Ty (H) in amastigotes also localize to the SPC, labeled by αCP1. Scale bars: 2 μm.