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. 2015 May 14;11(5):e1005233. doi: 10.1371/journal.pgen.1005233

Fig 2. 4E-BPs associate with translating ribosomes in an eIF4E-independent manner.

Fig 2

A. Sucrose density gradient analysis of extracts from the Eap1-TAP strain. Fractions were collected and protein distribution across the gradients visualised with western blots. Blots were probed for the 4E-BPs, closed loop components and 40S/60S ribosomal subunits. B. Western blot analyses of fractions collected from sucrose density gradients of extracts from the Eap1-TAP strain. The 80S ribosomal complex was dissociated by removal of cycloheximide and MgCl2 from the lysis buffer. Blots were probed for the 4E-BPs, closed loop components and 40S/60S ribosomal subunits. C. Western blot analysis of fractions collected from a sucrose density gradient of extract from the BY4741 HIS3 strain GP6001. The lysate was pre-treated with RNase I. D. Western blot analysis of fractions collected from a sucrose density gradient of extracts from the BY4741 HIS3 strain which had been grown in SCD-HIS and resuspended in SC-HIS for 10 min prior to cycloheximide treatment. E. Western blot analysis of FLAG immune-purification of whole cell extracts from caf20Δ strains harbouring an empty vector [pRS426], p[CAF20-FLAG] or p[CAF20 m2-FLAG]. F. Western blot analysis of fractions collected from sucrose density gradients of extracts from the p[CAF20-FLAG] and p[CAF20 m2-FLAG] strains.