Fig. 4: mTORC2-PKC-Nrf2 axis increases ATF4 abundance. | Cell Death Discovery

Fig. 4: mTORC2-PKC-Nrf2 axis increases ATF4 abundance.

From: Elucidation of an mTORC2-PKC-NRF2 pathway that sustains the ATF4 stress response and identification of Sirt5 as a key ATF4 effector

Fig. 4

A Western blot analysis of WCL from MDA-MB-231 and BT549 cells ± CB-839 (1 μM) for 24 h. Blots are representative of two (MDA-MB-231) or three (BT549) independent experiments. B Western blot analysis of WCL from MDA-MB-231 and BT549 cells treated with CB-839 (1 μM) ± Torin1 (500 nM) for 24 h. Blots are representative of three independent experiments. C Western blot analysis of WCL from control or sin1 KD MDA-MB-231 cells treated with CB-839 (1 μM) or deprived of glutamine for 24 h. Blots are representative of two independent experiments. D Western blot analysis of WCL from MDA-MB-231 and BT549 cells treated with CB-839 (1 μM) ± Ro31-8220 (5 μM) for 24 h. Blots are representative of two independent experiments. E Western blot analysis of WCL from control or PKCα KD MDA-MB-231 cells treated with CB-839 (1 μM) for 24 h. Blots are representative of two independent experiments. F Western blot analysis of WCL from MDA-MB-231 cells treated with CB-839 (1 μM) ± AI-1 (10 mM) for 12 h. Blots are representative of two independent experiments. G RT-qPCR quantification of ATF4 mRNA in MDA-MB-231 cells treated with CB-839 (1 μM) ± AI-1 (10 mM) for 24 h. Data shown are representative of two independent experiments and expressed as means ± SD for triplicate measurements. H, I RT-qPCR quantification of ATF4 mRNA (H) and Western blot analysis of WCL (I) in control and Nrf2 KD MDA-MB-231 and BT549 cells treated with CB-839 (1 μM) for 24 h. Data shown (I) are representative of two independent experiments and expressed as means ± SD for triplicate measurements. Blots are representative of three (MDA-MB-231) or two (BT549) independent experiments.

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