Journal: The Journal of Biological Chemistry

Article Title: Domain Mapping of Heat Shock Protein 70 Reveals That Glutamic Acid 446 and Arginine 447 Are Critical for Regulating Superoxide Dismutase 2 Function *

doi: 10.1074/jbc.M116.756122

Figure Lengend Snippet: Determination of amino acid residue(s) in hsp70 that is/are critical for SOD2 interactions. A, scatterplots of SOD2 activity following site-directed mutagenesis of the GERAMT-binding site residues. Each amino acid in the GERAMT sequence was replaced stepwise with alanine (A) or more conservative residues, glutamine and asparagine (B). The mutated peptides were incubated individually with aliquots of cell lysates, and SOD2 activity was measured by colorimetric assays (**, p < 0.05 from HBSS-treated controls (CTL), n = 4).

Article Snippet: SOD2-specific activity was measured using a colorimetric assay kit (706002, Cayman Chemical, Ann Arbor, MI) as described previously ( 2 , 3 ).

Techniques: Residue, Activity Assay, Mutagenesis, Binding Assay, Sequencing, Incubation

Journal: Mediators of inflammation

Article Title: The AMPK-mTOR Pathway Is Inhibited by Chaihu Shugan Powder, Which Relieves Nonalcoholic Steatohepatitis by Suppressing Autophagic Ferroptosis.

doi: 10.1155/2024/4777789

Figure Lengend Snippet: FIGURE 5: The progression of AMPK/mTOR-mediated autophagy and ferroptosis in MCD-fed mice can be mitigated by treatment with CSP, SSa, and NGN. (a) Observations of ultrathin liver sections were conducted using TEM (magnifications of 9700X and 23,000X). (b and c) ROS staining and relative expression levels (with a scale bar of 50 µm). (d–g) Liver Fe2+, SOD, MDA, and GSH levels. (h and i) Western blot

Article Snippet: Ferrous Iron (Fe2+) Colorimetric Assay Kit was acquired from Elabscience Biotechnology Co., Ltd., Wuhan, China.

Techniques: Staining, Expressing, Western Blot

Journal: Mediators of inflammation

Article Title: The AMPK-mTOR Pathway Is Inhibited by Chaihu Shugan Powder, Which Relieves Nonalcoholic Steatohepatitis by Suppressing Autophagic Ferroptosis.

doi: 10.1155/2024/4777789

Figure Lengend Snippet: FIGURE 6: In the NASH model, 3-MA administration had beneficial effects. (a–c) Liver and histological images of liver tissue sections stained with HE and Oil Red O, respectively. The scale bar measures 100 μm; the original magnification was 200 times. (d) Examination of ultrathin hepatic sections via transmission electron microscopy at magnifications of 9700X and 23,000X. (e) Liver weights across the various mouse cohorts. (f and g) Serum AST and ALT levels. (h–j) Levels of IL-1β, IL-6, and TNF-α in the liver. (k–n) Liver Fe2+, SOD, MDA, and GSH levels. (o) Western blot analysis confirmed the protein expression levels of KEAP1, Nrf-2, FTH1, and GPX4 in the three sample groups. ALT, alanine aminotransferase; AST, aspartate aminotransferase; GPX4, glutathione peroxidase 4; GSH, glutathione; HE, hematoxylin–eosin; IL, interleukin; KEAP1, Kelch-like ECH-associated protein 1; MDA, malondialdehyde; NASH, nonalcoholic steatohepatitis; Nrf-2, nuclear factor erythroid 2-related factor 2; SOD, superoxide dismutase; TNF-α, tumor necrosis factor-α. ∗p<0:05, ∗∗p<0:01, ∗∗∗p<0:001, and ∗∗∗∗p<0:0001.

Article Snippet: Ferrous Iron (Fe2+) Colorimetric Assay Kit was acquired from Elabscience Biotechnology Co., Ltd., Wuhan, China.

Techniques: Staining, Transmission Assay, Electron Microscopy, Western Blot, Expressing