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Figure 3 | Orphanet Journal of Rare Diseases

Figure 3

From: Rescue of nonsense mutations by amlexanox in human cells

Figure 3

Amlexanox is not toxic, does not inhibit general translation and does not affect natural NMD substrates expression. (A) Calu-6 Cell viability was measured after 20 hours of cell incubation with increasing amounts of amlexanox or with DMSO. These results combine three independent experiments. (B) Measure of translation efficiency after amlexanox treatment. Calu-6 Cells were incubated with DMSO, amlexanox or cycloheximide (CHX) before to measure translation efficiency using Click-iT AHA for nascent protein synthesis kit (Life technologies). This result is representative of 2 independent experiments. (C) Western-blot showing that the level of phosphorylated (Phos.) eIF2α does not increase in the presence of amlexanox in DMD cells (upper panels) or 6CFSMEo- cells (lower panels). The three left lanes represent a two-fold serial dilution of cell extract from untreated WT myoblasts (upper panels) or Calu-3 cells (lower panels). (D) The level of three NMD natural target mRNAs was measured from Calu-6 cells as described in Materials and Methods from three independent experiments.

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