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. 2018 Jul 2;217(7):2503–2518. doi: 10.1083/jcb.201711037

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© 2018 Biedka et al.

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Figure 3. — Dramatic changes in pre-60S subunit protein composition are caused by depletion of AF Nsa2. (A–C) Preribosomes containing (gal) or lacking (glu) Nsa2 were purified using TAP-tagged Nop7 as bait. (A) The protein composition of the pre-60S subunits was analyzed by SDS-PAGE followed by silver staining and Western blotting. Rsa4 is indicated by a black arrowhead; the contaminating band is IgG. (B and C) Semiquantitative mass spectrometry (iTRAQ) was used to quantify the relative changes in levels of 60S subunit r-proteins (B) and AFs (C) in the presence and absence of Nsa2. The ratios were normalized to levels of Nop7, and the fold change in log2 scale is shown for two biological replicates. A complete dataset for AFs is shown in Fig. S2.