The c.-201_-198delinsCTTT variant allele exhibits reduced translation potential in MCF7 cells treated with Torin1.

<p>The c.-201_-198delinsCTTT variant allele exhibits reduced translation potential in MCF7 cells treated with Torin1.</p

Functional analysis of a CDKN2A 5’UTR germline variant associated with pancreatic cancer development - Fig 5

<p>Relative p16 ^INK4a mRNA and protein expression in c.-201_-198delinsCTTT variant, measured by qPCR (A) and Western Blot (B); for qPCR, the average normalized expression of p16^INK4a and the standard deviation of three replicates are presented. (C) Polysomal c.-201_-198delinsCTTT allelic imbalance. Cytoplasmic lysates of the lymphoblastoid cell line heterozygous for the c.-201_-198delinsCTTT variant were fractionated using 15–50% sucrose gradients and RNAs co-sedimenting with subpolysomal fractions or with polysomes were extracted and used to amplify and sequence the p16^INK4a 5’UTR by Ion Torrent. The Y-axis on the left displays the allelic coverage, while the one on the right plots the percentage of relative imbalance of the wt allele in the comparison between polysomal and subpolysomal fractions. SUB = subpolysomal; POL = polysomal; Var = c.-201_-198delinsCTTT variant.</p

No impact of the <i>CDKN2A</i> 5’UTR c.-201_-198delinsCTTT variant was observed in the pGL3-promoter and in the bicistronic pRuf reporter vector as compared to the wt.

<p>The assay was performed in the pancreatic cell line BXPC-3 null for p16 ^INK4a protein.</p

The c.-201_-198delinsCTTT variant allele exhibits reduced translation potential in MCF7 cells treated with Torin1.

<p>The c.-201_-198delinsCTTT variant allele exhibits reduced translation potential in MCF7 cells treated with Torin1.</p