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. 2017 Apr;23(4):567–577. doi: 10.1261/rna.060236.116

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© 2017 Friesen et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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FIGURE 6. — Clitocine induces readthrough of p53 premature stop codons to produce full-length, functional p53 protein. (A) Western blot analysis showing dose response and time course of full-length p53 production after clitocine treatment of cell lines (from left to right) with native p53 nonsense alleles (CAOV-3_p53-136UAA, CALU-6_p53-196UGA, and H520_p53-146UGA) and with wild-type p53 (293H). (B) Western blot (p53) and (C) graph of relative p21^waf1 mRNA levels measured by RT-qPCR showing that in H520_p53-146UGA cells, clitocine increases p53 protein which, in turn, transactivates p21^waf1mRNA. pFC-p53 indicates transient transfection of a plasmid that expresses p53 protein. Error bars represent standard deviation of the mean for three determinations. (D) Caspases 3 and 7 activity and (E) cell viability assays showing that cells containing p53 premature stop codon alleles are more sensitive to the activity of clitocine. Error bars represent standard deviation of the mean for three determinations. (F) Results of a xenograft study showing that clitocine at doses as low as 0.3 mg/kg (five times per week) reduced the growth of CAOV-3_p53-UAA136 tumors. Error bars represent standard error of the mean for 10 mice per group.