Fig. 5: Glucose deprivation transactivates DCAF1, promotes ubiquition-mediated degradation of Rheb, and inhibits mTORC1 activity. | Cell Death & Disease

Fig. 5: Glucose deprivation transactivates DCAF1, promotes ubiquition-mediated degradation of Rheb, and inhibits mTORC1 activity.

From: Glucose deprivation triggers DCAF1-mediated inactivation of Rheb-mTORC1 and promotes cancer cell survival

Fig. 5

A, B Silencing DCAF1 delays glucose deprivation-induced Rheb degradation and enhances the protein stability of Rheb. Stable sgDCAF1 and sgControl cells were cultured in glucose-free medium, treated without (A) or with (B) 100 µg/ml CHX, and collected at indicated times for IB analysis. C Stable sgDCAF1 cells were cultured with or without glucose-free medium, and the polyubiquitination of Rheb was analyzed by immunoprecipitation. D Huh7, HCT116, and EC109 cells were cultured in glucose-free medium and cell protein was collected at indicated times for IB analysis. E Huh7, HCT116, and EC109 cells were cultured in glucose-free medium and total RNA was collected at indicated times. The mRNA levels of DCAF1 were analyzed using quantitative-PCR (Q-PCR). F Cells were treated with 20 mM 2-DG, and total RNA was collected at indicated times. The DCAF1 mRNA levels were analyzed using Q-PCR. G, H Effect of 2-DG treatment on the protein level of DCAF1, Rheb, and mTORC1 activity in mouse model. The 6-week-old C57BL6 mice were randomized into two groups and treated with normal saline or 2-DG solution for 3 weeks. The liver (G) and brain (H) tissues were harvested, and protein expression was evaluated by IB analysis using specific antibodies as indicated. All data were representative of at least three independent experiments (n = 3).

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