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Fig. 1 | BMC Biology

Fig. 1

From: Inhibiting eukaryotic ribosome biogenesis

Fig. 1

Screen setup to identify novel ribosome biogenesis inhibitors. a Reporter proteins for either the 40S (Rps9a) or the 60S (Rpl7a) ribosomal subunit were C-terminally fused to GFP. The resulting reporter strains were separately tested with ~ 1000 substances from two compound libraries (NIH clinical collection (Additional file 2: Table S1) and Screen-Well Natural Product Library Version 7.4 (Enzo Life sciences) (Additional file 2: Table S2)). b After treatment, cells were inspected by fluorescence microscopy. Ribosome biogenesis defects were identified by a shift of the steady-state GFP signal of the reporters from the cytoplasm into the nucleolus (NL) and/or the nucleoplasm (NP). c A total pool of 128 positively scoring inhibitors comprised 16 inhibitors specific for the 60S subunit, 96 specific for the 40S subunit and 16 affecting both subunits. d Positively scoring hits were further characterized by northern blot analysis of pre-rRNAs and subsequent hierarchical clustering based on quantification of processing intermediates

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