Journal: RNA Biology

Article Title: Interplay between Y-box-binding protein 1 (YB-1) and poly(A) binding protein (PABP) in specific regulation of YB-1 mRNA translation

doi: 10.4161/rna.8.5.16022

Figure Lengend Snippet: YB-1 and PABP effects on translation of YB-1 mRNA and its mutant forms. (A and C) 0.1 pmol of C+A− wild type, mutated YB-1 mRNAs, and Luciferase mRNA, as internal control, were translated in a rabbit reticulocyte cell-free system in the presence of increasing amounts of YB-1 (3.5 pmol, lanes 2, 7, 12, 17, 22; 7 pmol, lanes 3, 8, 13, 18, 23; 10.5 pmol, lanes 4, 9, 14, 19, 24; 14 pmol, lanes 5, 10, 15, 20, 25) or without it (lanes 1, 6, 11, 16, 21) (A) and in the presence of increasing amounts of PABP (2 pmol, lanes 2, 6, 10, 14, 18; 4 pmol, lanes 3, 7, 11, 15, 19; 6 pmol, lanes 4, 8, 12, 16, 20) or without it (lanes 1, 5, 9, 13, 17) (C). [14C]Tyr-labeled translation products were resolved by SDS-PAGE and visualized by autoradiography. (B and D) The relative amount of radioactivity in the bands of (A and C), respectively, was determined using a Packard Cyclone Storage Phosphor System (Packard Instrument Company, Inc.). The level of translation of each mRNA without addition of YB-1 or PABP was taken to be 100%. Values are the means of at least three independent experiments. Errors are standard deviations. Student test was used to confirm the statistic significance of differences between values for WT and mutant YB-1 mRNAs. ***p < 0.001; **p < 0.01; *p < 0.05.

Article Snippet: [ 14 C]Tyr-labeled translation products were resolved by SDS-PAGE and visualized by autoradiography. (B and D) The relative amount of radioactivity in the bands of (A and C), respectively, was determined using a Packard Cyclone Storage Phosphor System (Packard Instrument Company, Inc.).

Techniques: Mutagenesis, Luciferase, Control, Labeling, SDS Page, Autoradiography, Radioactivity