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. 2019 Nov 26;20(23):5941. doi: 10.3390/ijms20235941

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Mutation of either Puf6, Loc1, or Rpl43 showed similar rRNA processing defects. (A) Pre-90S and pre-60S ribosomal subunits at different stages were immunoprecipitated and detected by Western blotting. (B) The rRNA processing pathway and the probes used in the Northern blotting. (C,D) The rRNA processing was detected in the strains below with Northern blotting. (C) Wildtype (WT) and GAL::RPL43 strains were cultured in the medium containing galactose. Glucose was added to stop the transcription of the GAL-driven promoter to shut down the expression of Rpl43. (D) WT, puf6Δ (KLY67), loc1Δ (KLY218), and puf6Δloc1Δ (KLY312) were cultured at 30 °C until OD = 0.4~0.6. The cells were collected for RNA extraction. (C,D) The quantifications are shown below each sample. The relative intensities compared to initial state (C) and WT strain (D) were calculated.