ZFAS1 is associated with actively translating ribosomes. (A) Polysome distribution of MDA-MB-468 cell lysates as separated on a 7–47 % sucrose gradient. Absorbance at 260 nm is shown on the Y axis. Fractions from the top of the gradient to the bottom are shown from left to right on the X axis. Fractions were collected in 36 equal volumes, of which every third was used for RNA extraction, and cDNA synthesised for PCR. (B) Polysome distribution of MDA-MB-468 cell lysate separated on a 7–47 % sucrose gradient containing EDTA instead of MgCl2. Loss of the polysome peak is observed, together with a leftward shift of the ribosome subunits. (C) and (D) Quantitative expression of ZFAS1 and GAPDH measured by qPCR relative to 18S and 28S rRNAs prepared with and without the addition of EDTA. Arrows indicate where ribosomal features are observed on profiles in relation to fraction number. (PDF 536 kb
