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. 2017 Feb 16;65(4):743–750.e4. doi: 10.1016/j.molcel.2016.11.039

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© 2017 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

Ubiquitination of eS10 Facilitates Initiation of the RQC Pathway

(A) Cytosol from cells stably expressing different HA-tagged variants of eS10 was analyzed by immunoblotting for eS10 and the HA tag.

(B) Cytosol from cells stably expressing the HA-tagged K138/139R mutant of eS10 was separated on a 10%–50% sucrose gradient and immunoblotted for eS10. The fractions containing ribosomes are indicated. Similar results were seen for other eS10 variants (not shown).

(C) The indicated HA-eS10 expressing cells were transfected with the (K^AAA)₂₀ reporter and analyzed by flow cytometry 24 hr later. The RFP:GFP ratio of all transfected cells is shown as a histogram (eS10-HA in gray, eS10-K139R-HA in red, eS10-K138/139R-HA in blue). See also Figure S4.