Figure 2 from Combining Brigatinib with mTOR Inhibition to Effectively Treat NF2-SWN–Associated and Sporadic NF2-Deficient Meningiomas

Immortalized AG-NF2-Men cells demonstrated robust telomerase activity. A, Real-time PCR amplification curves of a telomerase-specific template after incubation with cell extracts from primary AG-NF2-Men cells and telomerase-immortalized AG-NF2-Men and Ben-Men-1 cells. Manufacturer-supplied lysates from telomerase-expressing cells before and after heat inactivation served as positive and negative assay controls, respectively. A no-template control (H2O) was used as an additional negative control. The horizontal red line indicates the threshold at which the Ct values were measured. B and C, Melt curve (B) and Melt peak (C) analysis of the telomeric repeat amplicons at the end of the PCR analysis. D, Tables summarizing the mean Ct and melt temperatures of the amplicons and the estimated fold increase in telomerase activity in immortalized AG-NF2-Men and Ben-Men-1 cells vs. primary AG-NF2-Men cells. The telomerase activity in immortalized AG-NF2-Men cells was ∼6.6 × 104-fold higher than that in the primary culture. E, An agarose gel showing that high levels of telomerase-mediated elongation of the synthetic telomeric repeat template occurs only in telomerase-immortalized AG-NF2-Men and Ben-Men-1 cells.