Neuroendocrine marker expression was increased by BYL719 treatment.

<p>A: BON-1, H727 and QGP-1 cells were treated with 10 μM BYL719 for 96 h and mRNA expression of Chromogranin A <i>(CHGA)</i>, <i>SSTR1</i>, <i>SSTR2</i> and <i>SSTR5</i> was analyzed by qPCR. B: BON-1, H727 and QGP-1 cells were treated with 1 μM BYL719 for 96 h and mRNA expression of <i>SSTR2</i> was analyzed by qPCR; Values are summarized from three independent experiments of three replicates per data point. * p<0.05; ** p≤0.01; *** p≤0.001.</p

Assessment of apoptosis: Caspase 3/7 assay results after 24 h treatment with different doses of BYL719 are shown as mean percentage of caspase 3/7 activity referred to the untreated control (100%) ± SD: Strongest apoptosis was induced in BON-1 cells, followed by H727 cells.

<p>QGP-1 cells showed the least but still significant apoptosis. * p<0.05; ** p≤0.01; *** p≤0.001 (two independent experiments with three replicates per data point).</p

The combination therapy with BYL719 plus everolimus strongly induced SSTR2 expression in BON-1 and QGP-1 cells.

<p>Cells have been treated with 5 μM BYL719, 5 nM everolimus, or the combination of both for 48h (A) or with 1 μM BYL719, 1 nM everolimus or the combination of both for 96h (B). SSTR2 expression was analyzed by Western Blot (A) and qPCR (B). Representative western blot of two replicates; qPCR results are summarized from three independent experiments of three replicates. * p<0.05; ** p≤0.01; *** p≤0.001.</p