Journal: Frontiers in Neuroscience

Article Title: Isolation and monoculture of functional primary astrocytes from the adult mouse spinal cord

doi: 10.3389/fnins.2024.1367473

Figure Lengend Snippet: Products used for functional assessments.

Article Snippet: xCT antibody-BSA Free , NB300-317 , Novus , Immunoblotting , 1:1000.

Techniques: Functional Assay, Western Blot, Concentration Assay, cDNA Synthesis, SYBR Green Assay, Saline

Journal: Pharmacological research

Article Title: Nrf2 and its dependent autophagy activation cooperatively counteract ferroptosis to alleviate acute liver injury.

doi: 10.1016/j.phrs.2022.106563

Figure Lengend Snippet: Fig. 1. SFN treatment activates Nrf2 and autophagy and reduces ferroptosis induced by H2O2 in hepatocytes. L02 and BRL cells were treated with H2O2 and SFN for 24 h. ROS (A), GSH/GSSG ratio (B), iron content (C), caspase-3 (D) and lipid ROS (I) were assessed to reflect the protective effect of SFN treatment on cells. Western blotting was used to detect the protein expression of Nrf2, SLC7A11, GPX-4, P62, and LC3II in L02 cells (E, F) and BRL cells (G, H). A ChIP assay was used to detect the binding of Nrf2 to the Atg5 promoter region (J). The expression of Nrf2, P62, and LC3II in L02 cells after overexpression of Nrf2 were also detected by western blotting (K). Data are expressed as mean ± SD (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: Antibodies against SLC7A11 (NB100–59735) were purchased from Novus Biologicals Co. (Colorado, USA).

Techniques: Western Blot, Expressing, Binding Assay, Over Expression

Journal: Pharmacological research

Article Title: Nrf2 and its dependent autophagy activation cooperatively counteract ferroptosis to alleviate acute liver injury.

doi: 10.1016/j.phrs.2022.106563

Figure Lengend Snippet: Fig. 3. Inhibition of Nrf2 attenuates SFN-induced autophagy and anti-ferroptosis ability in hepatocytes. Western blotting was used to detect the protein expression of Nrf2, SLC7A11, GPX-4, P62, and LC3II in L02 cells (A, B) and BRL cells (C, D) after ML385 intervention as well as their expression in L02 cells (E, F) after siNrf2 intervention. Data are expressed as mean ± SD (n = 3). * P < 0.05, * * P < 0.01, * ** P < 0.001, * ** * P < 0.0001.

Article Snippet: Antibodies against SLC7A11 (NB100–59735) were purchased from Novus Biologicals Co. (Colorado, USA).

Techniques: Inhibition, Western Blot, Expressing

Journal: Pharmacological research

Article Title: Nrf2 and its dependent autophagy activation cooperatively counteract ferroptosis to alleviate acute liver injury.

doi: 10.1016/j.phrs.2022.106563

Figure Lengend Snippet: Fig. 4. Inhibition of autophagy decreases the membrane transfer of SLC7A11 and weakens the protective effect of SFN in hepatocytes. Western blotting was used to detect the protein expression of Nrf2, SLC7A11, GPX-4, P62, and LC3II in L02 cells (A, B) and BRL cells (C, D). The expression of SLC7A11 in the membrane and cytoplasm of L02 cells (E, F) and BRL cells (G, H) were also detected by western blotting. Data are expressed as mean ± SD (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: Antibodies against SLC7A11 (NB100–59735) were purchased from Novus Biologicals Co. (Colorado, USA).

Techniques: Inhibition, Membrane, Western Blot, Expressing

Journal: Pharmacological research

Article Title: Nrf2 and its dependent autophagy activation cooperatively counteract ferroptosis to alleviate acute liver injury.

doi: 10.1016/j.phrs.2022.106563

Figure Lengend Snippet: Fig. 5. Inhibition of autophagy promotes colocalization of SLC7A11 with ly sosomes in L02 cells. Immunofluorescence was used to demonstrate the colocalization of SLC7A11 and Lamp2 in L02 cells.

Article Snippet: Antibodies against SLC7A11 (NB100–59735) were purchased from Novus Biologicals Co. (Colorado, USA).

Techniques: Inhibition, Immunofluorescence

Journal: Pharmacological research

Article Title: Nrf2 and its dependent autophagy activation cooperatively counteract ferroptosis to alleviate acute liver injury.

doi: 10.1016/j.phrs.2022.106563

Figure Lengend Snippet: Fig. 6. SFN pretreatment alleviates ferroptosis and CCL4-induced liver injury in rats. Schematic of the in vivo study design (A). Biochemical indexes in serum (ALT, AST) (B). Biochemical indexes in the liver (ROS, GSH/GSSG ratio, iron content, and caspase-3) (C, D). Images of HE-stained of rat liver (E). Images of TUNEL-stained rat liver (F). TEM was used to detect mitochondrial morphology (G). Western blotting was used to detect the protein expression of Nrf2, SLC7A11 GPX-4, P62, and LC3II in rat liver (H). Data are expressed as mean ± SD (n = 5). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: Antibodies against SLC7A11 (NB100–59735) were purchased from Novus Biologicals Co. (Colorado, USA).

Techniques: In Vivo, Staining, TUNEL Assay, Western Blot, Expressing

Journal: Pharmacological research

Article Title: Nrf2 and its dependent autophagy activation cooperatively counteract ferroptosis to alleviate acute liver injury.

doi: 10.1016/j.phrs.2022.106563

Figure Lengend Snippet: Fig. 7. Nrf2 knockdown significantly attenuates SFN-induced autophagy and its hepatoprotective effect in rats. Images of HE-stained rat liver (A). Western blotting was used to detect the protein expression of Nrf2, SLC7A11, GPX-4, P62, and LC3II in rat liver (B). Data are expressed as mean ± SD (n = 5). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: Antibodies against SLC7A11 (NB100–59735) were purchased from Novus Biologicals Co. (Colorado, USA).

Techniques: Knockdown, Staining, Western Blot, Expressing

Journal: Pharmacological research

Article Title: Nrf2 and its dependent autophagy activation cooperatively counteract ferroptosis to alleviate acute liver injury.

doi: 10.1016/j.phrs.2022.106563

Figure Lengend Snippet: Fig. 9. Inhibition of autophagy increases the colocalization of SLC7A11 with lysosomes in rats liver tissue. Immunofluorescence was used to reflect the colocalization of SLC7A11 and Lamp2 in rat liver tissues.

Article Snippet: Antibodies against SLC7A11 (NB100–59735) were purchased from Novus Biologicals Co. (Colorado, USA).

Techniques: Inhibition, Immunofluorescence

Journal: Pharmacological research

Article Title: Nrf2 and its dependent autophagy activation cooperatively counteract ferroptosis to alleviate acute liver injury.

doi: 10.1016/j.phrs.2022.106563

Figure Lengend Snippet: Fig. 8. Inhibition of autophagy reduces the membrane transfer of SLC7A11 and attenuates the hepatoprotective effect of SFN in rats. Images of HE-stained rat liver (A). Western blotting was used to detect the protein expression of Nrf2, SLC7A11, GPX-4, P62, and LC3II in rat liver (B), and the expression of SLC7A11 in the cell membrane and cytoplasm in rat liver (C). Data are expressed as mean ± SD (n = 5). P < 0.05, * * P < 0.01, * ** P < 0.001, * ** * P < 0.0001.

Article Snippet: Antibodies against SLC7A11 (NB100–59735) were purchased from Novus Biologicals Co. (Colorado, USA).

Techniques: Inhibition, Membrane, Staining, Western Blot, Expressing

Journal: Pharmacological research

Article Title: Nrf2 and its dependent autophagy activation cooperatively counteract ferroptosis to alleviate acute liver injury.

doi: 10.1016/j.phrs.2022.106563

Figure Lengend Snippet: Fig. 10. Inhibition of autophagy promotes BECN1 phosphorylation through the AMPK pathway and inhibits SLC7A11 membrane localization. Western blotting was used to detect the protein expression of P-AMPK, AMPK, P-BECN1, and BECN1 in L02 cells (A, B). IP analysis was used to detect the formation of P-BECN1–SLC7A11 in L02 cells (C, D). Data are expressed as mean ± SD (n = 3). P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: Antibodies against SLC7A11 (NB100–59735) were purchased from Novus Biologicals Co. (Colorado, USA).

Techniques: Inhibition, Phospho-proteomics, Membrane, Western Blot, Expressing