Fig. 7: Evidence for the mTORC2-PKC-Nrf2-ATF4-Sirt5 pathway in brain cancer.

A Scatter plots showing that Sirt5 mRNA levels correlate with sin1 (Spearman: 0.27 Pearson: 0.96), PKCA (Spearman: 0.37 Pearson: 0.92), NRF2 (Spearman: 0.23 Pearson: 0.96) and ATF4 (Spearman: 0.38 Pearson: 0.97) mRNA levels in brain cancer. Data are from the TGCA brain cancer dataset. B RT-qPCR quantification of ATF4 mRNA in LN229 cells cultured ± glutamine in serum free medium for 24 h. Data shown are representative of two independent experiments and expressed as means ± SD for triplicate measurements. C RT-qPCR quantification of Sirt5 mRNA in LN229 cells cultured ± glutamine in serum free medium for 24 h. Data shown are representative of two independent experiments and expressed as means ± SD for triplicate measurements. D RT-qPCR quantification of Sirt5 mRNA in control and ATF4 KD LN229 cells cultured in glutamine free medium for 24 h. Data shown are representative of two independent experiments and expressed as means ± SD for triplicate measurements. E Western blot analysis of WCL from LN229 cells treated with Torin1 (500 nM), Rapamycin (50 nM), MK2206 (5 μM), Ro31-8220 (1 μM), AI-1 (10 mM) or ActD (1 μg/mL) in serum/glutamine free medium for 24 h. Blots are representative of two independent experiments.