Journal: Toxins

Article Title: Activation of the IRE1α Arm, but not the PERK Arm, of the Unfolded Protein Response Contributes to Fumonisin B1-Induced Hepatotoxicity

doi: 10.3390/toxins12010055

Figure Lengend Snippet: Activation of the IRE1α axis, but not PERK axis, of the ER stress response contributed to FB1-induced ER stress-mediated hepatocyte toxicity. AML12 cells were treated with 50 to 300 μM FB1 for 48 h, and then ( A ) the inhibitory effects of FB1 on the AML12 cells was evaluated by crystal violet staining, and ( B ) the cell death was analyzed by Annexin V/PI staining. ( C ) The effect of FB1 on the ER stress makers, the cells were exposed to FB1 for 48 h, and the phosphorylation of PERK, IRE1α, and eIF2α were analyzed by Western blot. n = 3 ( D ) The influence of the ER stress inhibitor TUDCA on the cell death induction by FB1. ( E & F ) The effect of IRE1α or PERK specific inhibitor on FB1-induced cell death in AML12 cells and MEF cells. The cells were exposed to FB1 in the presence or absence of inhibitors for the indicated time, and the cell death was evaluated by the flow cytometry analysis of Annexin V/PI positive cells. The bars denote standard errors from three experiments. * p < 0.05, ** p < 0.01, *** p < 0.001 compared with the corresponding control.

Article Snippet: IRE1α inhibitor 4μ8C and PERK inhibitor GSK2606414 were purchased from MCE (Shanghai, China).

Techniques: Activation Assay, Staining, Phospho-proteomics, Western Blot, Flow Cytometry, Control

Journal: Toxins

Article Title: Activation of the IRE1α Arm, but not the PERK Arm, of the Unfolded Protein Response Contributes to Fumonisin B1-Induced Hepatotoxicity

doi: 10.3390/toxins12010055

Figure Lengend Snippet: The IRE1α-mediated activation of the mitochondrial pathway plays an important role in apoptosis induction by FB1 in liver cells. ( A ) The effect of FB1 on the expression of JNK, Mcl-1, Bak, Bax, and Puma in the protein level. The cells were exposed to FB1 with or without TUDCA for 48 h, and the phosphorylation of JNK, Mcl-1, Bak, Bax, and Puma were analyzed by Western blotting. n = 3. ( B ) FB1 significantly induced cell death in wild-type MEF cells but not in Bax/Bak knockout MEF cells. The bars denote standard errors from three experiments. ( C ) The effect of the IRE1α specific inhibitor 4μ8C on the expression of apoptosis-related proteins. The cells were exposed to FB1 with or without 4μ8C for 24 h, and the phosphorylation of JNK, Mcl-1, Bak, Bax, and Puma were analyzed by Western blotting. n = 3. ** p < 0.01 compared with the corresponding control.

Article Snippet: IRE1α inhibitor 4μ8C and PERK inhibitor GSK2606414 were purchased from MCE (Shanghai, China).

Techniques: Activation Assay, Expressing, Phospho-proteomics, Western Blot, Knock-Out, Control

Journal: Toxins

Article Title: Activation of the IRE1α Arm, but not the PERK Arm, of the Unfolded Protein Response Contributes to Fumonisin B1-Induced Hepatotoxicity

doi: 10.3390/toxins12010055

Figure Lengend Snippet: ROS generation is responsible for FB1-induced ER stress. ( A ) The effects of FB1 on the ROS generation. The cells were treated with FB1 for 8 h or 18 h, and the intercellular ROS levels were assessed using flow cytometry after DCFH-DA staining. ( B ) The influence of ROS suppression by NAC on the ER stress markers and cell apoptosis. The cells were treated with FB1 with or without NAC for 48 h, and the phosphorylation of IRE1α and eIF2α, Bip, and cleaved PARP were analyzed by Western blotting. n = 3. ( C ) The influence of ROS suppression by NAC on cell death induction. The cells were treated with FB1 with or without NAC for 48 h, and the cell death was evaluated by flow cytometry analysis after Annexin V staining. ( D ) The effect of the IRE1α or PERK specific inhibitor on FB1-induced ROS. The bars denote standard errors from three experiments. ** p < 0.01, *** p < 0.001 compared with the corresponding control.

Article Snippet: IRE1α inhibitor 4μ8C and PERK inhibitor GSK2606414 were purchased from MCE (Shanghai, China).

Techniques: Flow Cytometry, Staining, Phospho-proteomics, Western Blot, Control

Journal: Toxins

Article Title: Activation of the IRE1α Arm, but not the PERK Arm, of the Unfolded Protein Response Contributes to Fumonisin B1-Induced Hepatotoxicity

doi: 10.3390/toxins12010055

Figure Lengend Snippet: Glycyrol (GC) prevents FB1-induced apoptosis in AML12 cells through inactivating IRE1α. The effects of GC on FB1-induced cell death. ( A ) The cells were exposed to 300 μM FB1 with or without 10 μM GC for 48 h, and then the cells were collected for a death measurement by Annexin V/PI staining. The bars denote standard errors from three experiments. ( B ) The influences of GC on the FB1-mediated ER stress. The cells were exposed to 300 μM FB1 with or without 10 μM GC for 24 h, and then IRE1α phosphorylation and Bip were examined by Western blotting. n = 3. ** p < 0.01 compared with the corresponding control.

Article Snippet: IRE1α inhibitor 4μ8C and PERK inhibitor GSK2606414 were purchased from MCE (Shanghai, China).

Techniques: Staining, Phospho-proteomics, Western Blot, Control