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"To asses autophagy induction MAP1LC3B, GABARAPL2, and GABARAP levels were determined by western blot as indicated under Materials and Methods."
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"To asses autophagy induction MAP1LC3B, GABARAPL2, and GABARAP levels were determined by western blot as indicated under Materials and Methods."
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"To asses autophagy induction MAP1LC3B, GABARAPL2, and GABARAP levels were determined by western blot as indicated under Materials and Methods."
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"To asses autophagy induction MAP1LC3B, GABARAPL2, and GABARAP levels were determined by western blot as indicated under Materials and Methods."
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"(See previous page). SIRT5 controls ammonia-induced autophagy. (A) MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5+ and SIRT5− clones were processed to obtain whole cellular extracts. Alternatively, WT, SIRT5+ and SIRT5− cells were treated with 100 nM bafilomycinA1 for 2 and 17 h and processed. Finally, also MDA-MB-231 WT, SIRT3+ and SIRT3− cells were used. Autophagy induction was measured by using an anti-MAP1LC3B antibody as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. *Significantly different from WT cells. Significance was set atP < 0.05. (B) MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5+ and SIRT5− clones were processed to obtain whole cell extracts. SQSTM1, GABARAPL2, and GABARAP levels were determined by western blot as indicated under Materials and Methods. In addition, cells were treated with bafilomycinA1 for 17 h and SQSTM1 levels measured by western blot. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. *Significantly different from WT cells. Significance was set atP < 0.05. (C) MDA-MB-231 WT cells were either left untreated or treated with NH4Cl 1 or 2 mM for 24 h. Subsequently, cells were processed to obtain whole extracts. MAP1LC3B, GABARAPL2, GABARAP and, SQSTM1 levels were measured by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. *Significantly different from WT cells. Significance was set atP < 0.05. (D) MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5+ and SIRT5− clones were treated with either BPTES or dimethyl-α-ketoglutarate. Cells were then processed to obtain whole extracts. MAP1LC3B, GABARAPL2, and, SQSTM1 levels were measured by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. *Significantly different from WT cells. Significance was set atP < 0.05. (E) MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5+ and SIRT5− clones were cultured withoutL-glutamine for 24 h. Cells were then processed to obtain whole extracts. MAP1LC3B and SQSTM1 levels were measured by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. *Significantly different from WT cells. Significance was set atP < 0.05. (F) C2C12 WT cells treated in the presence or absence of MC3482, as well as SIRT5+ and SIRT5− clones were processed to obtain whole cell extracts. Cells were also treated with NH4Cl 1 or 2 mM for 24 h. To asses autophagy induction MAP1LC3B, GABARAPL2, and GABARAP levels were determined by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. *Significantly different from WT cells. Significance was set atP < 0.05.\n",
"Ammonia-induced autophagy is regulated by SIRT5",
"Results",
"SIRT5 regulation of ammonia-induced autophagy and mitophagy"
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"paragraph.text": [
"(See previous page). SIRT5 controls ammonia-induced autophagy. (A) MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5+ and SIRT5− clones were processed to obtain whole cellular extracts. Alternatively, WT, SIRT5+ and SIRT5− cells were treated with 100 nM bafilomycinA1 for 2 and 17 h and processed. Finally, also MDA-MB-231 WT, SIRT3+ and SIRT3− cells were used. Autophagy induction was measured by using an anti-MAP1LC3B antibody as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. *Significantly different from WT cells. Significance was set atP < 0.05. (B) MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5+ and SIRT5− clones were processed to obtain whole cell extracts. SQSTM1, GABARAPL2, and GABARAP levels were determined by western blot as indicated under Materials and Methods. In addition, cells were treated with bafilomycinA1 for 17 h and SQSTM1 levels measured by western blot. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. *Significantly different from WT cells. Significance was set atP < 0.05. (C) MDA-MB-231 WT cells were either left untreated or treated with NH4Cl 1 or 2 mM for 24 h. Subsequently, cells were processed to obtain whole extracts. MAP1LC3B, GABARAPL2, GABARAP and, SQSTM1 levels were measured by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. *Significantly different from WT cells. Significance was set atP < 0.05. (D) MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5+ and SIRT5− clones were treated with either BPTES or dimethyl-α-ketoglutarate. Cells were then processed to obtain whole extracts. MAP1LC3B, GABARAPL2, and, SQSTM1 levels were measured by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. *Significantly different from WT cells. Significance was set atP < 0.05. (E) MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5+ and SIRT5− clones were cultured withoutL-glutamine for 24 h. Cells were then processed to obtain whole extracts. MAP1LC3B and SQSTM1 levels were measured by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. *Significantly different from WT cells. Significance was set atP < 0.05. (F) C2C12 WT cells treated in the presence or absence of MC3482, as well as SIRT5+ and SIRT5− clones were processed to obtain whole cell extracts. Cells were also treated with NH4Cl 1 or 2 mM for 24 h. To asses autophagy induction MAP1LC3B, GABARAPL2, and GABARAP levels were determined by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. *Significantly different from WT cells. Significance was set atP < 0.05."
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