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Western blot of total cell lysates (TCL) and co-precipitated streptavidin-bound proteins (IP: Strep [biotin]) from WT and stm1Δ cells expressing endogenously tagged Rpl16a/b-HA-TEV-AVI. Samples were uninduced (−Estr) or with 5 nM estradiol for 2 h to induce BirA expression. After estradiol washout, samples were supplemented with biotin to induce labeling of Rpl16a/b-HA-TEV-AVI and 1 mM auxin (IAA) to initiate BirA degradation. Samples were harvested after the BirA biotinylation/degradation step (CHASE, 0 h) or 1 h after BirA shut-off and degradation (CHASE, 1 h). Blots were probed with HA antibodies (biotinylated Rpl16a/b-HA-TEV-AVI after TEV elution from Strep-beads), endogenous Rps5 and Rpl35, Stm1, and Pgk1 (loading control). N =3 independent replicates. Blue boxes = Rpl16a/b and Rpl35. Orange boxes = Rps5.

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