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. 2021 Oct 31;18(6):1416–1432. doi: 10.1080/15548627.2021.1987672

Figure 7.

Figure 7.

FAIM suppresses the interaction between ULK1 and MTOR in cytoplasm. (A) H1299 cells were lysed and immunoprecipitation was performed using the indicated antibodies. Western blot were used to detect protein expression. (B) H1299 cells were transfected with or without pcDNA3.1-HA-FAIM. Forty-eight h later, the cells were lysed and immunoprecipitation was performed. The expression of the indicated proteins was examined by western blot. (C) H1299 cells were transfected with pCMV-HA-ULK1 and the indicated siRNAs. Forty-eight h later, the cells were lysed and immunoprecipitation was performed. The expression of the indicated proteins was examined by western blot. (D) H1299 cells were co-transfected with pCMV-HA-ULK1 and pcDNA3.1-HA-FAIM. Forty-eight h later, the cells were lysed and immunoprecipitation was performed. The expression of the indicated proteins was examined by western blot. (E) A549 cells were transfected with pcDNA3.1-His-FAIM-ΔN. Forty-eight h later, Mitochondria were isolated and the mitochondria and cytosolic proteins were detected by western blot. (F) A549 cells were transfected with pcDNA3.1-His-FAIM-ΔN. Forty-eight h later, the cells were lysed and western blot was performed to detect the expression of the indicated proteins. (G) A549 cells were transfected with pcDNA3.1-His-FAIM-ΔN. Forty-eight h later, the cells were lysed and the production of α-ketoglutarate (α-KG) was measured. Data represent the average of three independent experiments (mean ± SD). ns, P > 0.05. (H) A549 cells were transfected with pcDNA3.1-His-FAIM-ΔN. Forty-eight h later, mitochondria were isolated and glutaminase activity was measured. Data represent the average of three independent experiments (mean ± SD). ns, P > 0.05. (I) A549 cells were transfected with or without pcDNA3.1-His-FAIM-ΔN. Forty-eight h later, the cells were lysed and immunoprecipitation was performed. The expression of the indicated proteins was examined by western blot. (J) A549 cells were co-transfected with pCMV-HA-ULK1 and pcDNA3.1-His-FAIM-ΔN. Forty-eight h later, the cells were lysed and immunoprecipitation was performed. The expression of the indicated proteins was examined by western blot.