Journal: Journal of Functional Foods

Article Title: Protective effects of selenium-enriched peptides from Cardamine violifolia on d-galactose-induced brain aging by alleviating oxidative stress, neuroinflammation, and neuron apoptosis

doi: 10.1016/j.jff.2020.104277

Figure Lengend Snippet: Fig. 7. CSP supplementation reduced D-gal-induced overexpression of RAGE, BACE-1, Aβ-42 and PS1. (A) Western blot analysis of RAGE, BACE-1, Aβ-42 and PS1; (B) The relative protein expressions of RAGE, BACE-1, Aβ-42 and PS1; Data are expressed as the mean ± SEM (n = 3). Differences were denoted as follows: * P < 0.05, ** P < 0.01, *** P < 0.001 compared with NC; # P < 0.05, ## p < 0.01, ### p < 0.001 compared with D-gal; & P < 0.05, && p < 0.01, &&& p < 0.001 compared with CSPL.

Article Snippet: Primary antibodies against NFkβ-p65, RAGE, BACE1, PS1, BAX, BCL2, Caspase-3, HO1, NQO1, and β-actin were purchased from Proteintech (Rosemont, IL, USA).

Techniques: Over Expression, Western Blot

Journal: Journal of Functional Foods

Article Title: Astragalin protects against lipopolysaccharide-triggered acute liver injury through suppression of necroptosis and inflammation and improvement of energy metabolism

doi: 10.1016/j.jff.2024.106298

Figure Lengend Snippet: Fig. 7. Astragalin (AST) eliminated lipopolysaccharide (LPS)-induced inflammatory response by inhibiting glycolysis enhancement mediated by the hypoxia- inducible factor-1α/pyruvate kinase M2 (HIF-1α/PKM2) signaling pathway. (A) LPS changed the concentrations of adenosine triphosphate (ATP) and lactic acid (LA) in liver and serum, and AST reversed these changes. Student’s t-test was performed, n = 4, * P < 0.05, ** P < 0.01, *** P < 0.001. (B) The activities of hexokinase (HK), pyruvate kinase (PK), and lactate dehydrogenase (LDH) in liver and serum were enhanced by LPS, whereas AST inhibited the enhancement. Student’s t-test was performed, n = 4, * P < 0.05, ** P < 0.01, *** P < 0.001. (C) Glycolysis-related genes, including HK2, LDHA, glucose-6-phosphate dehydrogenase X-linked (G6pdx), and glucose transporter 1 (Glut-1), were upregulated by LPS but AST prevented the upregulation. Similar results also showed in the expression of HIF-1α and PKM2 genes. Student’s t-test was performed, n = 4, * P < 0.05, ** P < 0.01, *** P < 0.001. (D-E) Expressions of HIF-1α and PKM2 proteins were increased by LPS treatment, while AST decreased their expression. Student’s t-test was performed, n = 3, * P < 0.05, ** P < 0.01.

Article Snippet: Adenosine triphosphate (ATP) content detection kit (#BC0305), lactic acid (LA) content assay kit (#BC2235), lactate dehydrogenase (LDH) activity assay kit (#BC0685), hexokinase (HK) activity assay kit (#BC0745), pyruvate kinase (PK) activity assay kit (#BC0545) were obtained from Beijing Solarbio Science & Technology Co., Ltd. Trizol (total RNA extraction reagent, #R0016), RIPA lysis buffer (strong, #P0013B), and BCA protein assay kit (#P0011) were provided by Beyotime Biotech Inc. Plus All-in-one 1st strand cDNA synthesis supermix (gDNA Purge, #E047-01B) and SYBR qPCR supermix plus (#E096-01B) were obtained from Novoprotein Scientific Inc. We got TNF-α polyclonal antibody (#17590–1-AP), TNFR1associated death domain protein (TRADD) polyclonal antibody (#15468–1-AP), HIF-1α polyclonal antibody (#20960–1-AP), PKM2, muscle2-specific monoclonal antibody (#60268–1-Ig), β-actin recombinant antibody (#81115–1-RR), and TANK-binding kinase 1 (TBK1) polyclonal antibody (#28397–1-AP) from Proteintech Group, Inc. Phospho-TBK1 (P-TBK1, Ser172) polyclonal antibody (#BD-PP1527) was provided by Biodragon Co. Ltd. Anti-phospho-RIP family of serinethreonine kinases (Ser166, P-RIPK1) polyclonal antibodies (#31122S), RIPK1 (D94C12) monoclonal antibody (#3493S), RIPK3 (D8J3L) monoclonal antibody (#15828S), and anti-phospho-RIPK3 (Thr231/ Ser232, E7S1R, #91702) were obtained from Cell Signaling Technology, Inc. Anti-phospho-MLKL (Ser345, P-MLKL) monoclonal antibody (#MABC1158) was provided by Merck Millipore Corporation, while MLKL polyclonal antibody (#PA5-71886) was provided by Thermo Fisher Scientific Inc. Bio-Rad Laboratories Co. Ltd provided CD68 monoclonal antibody (#MCA1957).

Techniques: Expressing

Journal: Journal of Functional Foods

Article Title: Astragalin protects against lipopolysaccharide-triggered acute liver injury through suppression of necroptosis and inflammation and improvement of energy metabolism

doi: 10.1016/j.jff.2024.106298

Figure Lengend Snippet: Fig. 8. The involved molecular mechanisms of astragalin (AST) in the treatment of lipopolysaccharide (LPS)-induced acute liver injury (ALI). LPS promotes the M1 type macrophage activation, causing inflammatory responses and the release of inflammatory cytokines, including tumor necrosis factor (TNF)-α. By binding to its receptor, TNF-α activated the receptor-interacting protein kinase 1 (RIPK1)/RIPK3/ mixed lineage kinase domain-like protein (MLKL) signal axis and mediated necroptosis. Meanwhile, LPS triggered the hypoxia-inducible factor-1α/pyruvate kinase M2 (HIF-1α/PKM2) signaling pathway, which mediates glycolysis-enhanced inflammatory response and oxidative stress. However, AST inhibited the transition of macrophages into M1 type, reduced the expression of pro-inflammatory cy tokines, which further decreased the activation of related signaling pathways, thereby preventing the liver from acute injury. This figure is created by Figdraw.

Article Snippet: Adenosine triphosphate (ATP) content detection kit (#BC0305), lactic acid (LA) content assay kit (#BC2235), lactate dehydrogenase (LDH) activity assay kit (#BC0685), hexokinase (HK) activity assay kit (#BC0745), pyruvate kinase (PK) activity assay kit (#BC0545) were obtained from Beijing Solarbio Science & Technology Co., Ltd. Trizol (total RNA extraction reagent, #R0016), RIPA lysis buffer (strong, #P0013B), and BCA protein assay kit (#P0011) were provided by Beyotime Biotech Inc. Plus All-in-one 1st strand cDNA synthesis supermix (gDNA Purge, #E047-01B) and SYBR qPCR supermix plus (#E096-01B) were obtained from Novoprotein Scientific Inc. We got TNF-α polyclonal antibody (#17590–1-AP), TNFR1associated death domain protein (TRADD) polyclonal antibody (#15468–1-AP), HIF-1α polyclonal antibody (#20960–1-AP), PKM2, muscle2-specific monoclonal antibody (#60268–1-Ig), β-actin recombinant antibody (#81115–1-RR), and TANK-binding kinase 1 (TBK1) polyclonal antibody (#28397–1-AP) from Proteintech Group, Inc. Phospho-TBK1 (P-TBK1, Ser172) polyclonal antibody (#BD-PP1527) was provided by Biodragon Co. Ltd. Anti-phospho-RIP family of serinethreonine kinases (Ser166, P-RIPK1) polyclonal antibodies (#31122S), RIPK1 (D94C12) monoclonal antibody (#3493S), RIPK3 (D8J3L) monoclonal antibody (#15828S), and anti-phospho-RIPK3 (Thr231/ Ser232, E7S1R, #91702) were obtained from Cell Signaling Technology, Inc. Anti-phospho-MLKL (Ser345, P-MLKL) monoclonal antibody (#MABC1158) was provided by Merck Millipore Corporation, while MLKL polyclonal antibody (#PA5-71886) was provided by Thermo Fisher Scientific Inc. Bio-Rad Laboratories Co. Ltd provided CD68 monoclonal antibody (#MCA1957).

Techniques: Activation Assay, Binding Assay, Expressing, Protein-Protein interactions

Journal: Cancer medicine

Article Title: The expression and role of SUZ12 in lung adenocarcinoma.

doi: 10.1002/cam4.70190

Figure Lengend Snippet: FIGURE 7 The effect of SUZ12 on CDKs and cyclins expression was tested by qRT-PCR and western blotting. sh-SUZ12 decreased CDK3 mRNA (A), CDK2/3 (C), and cyclin D1 (F) protein expression, while increased cyclin E1 protein expression (F), without significantly effected the cyclins mRNA (E). oe-SUZ12 increased CDK3/6 mRNA (B) and CDK3 protein expression (D). *p < 0.05, **p < 0.001.

Article Snippet: The list of primary antibodies: SUZ12 (1 μg/mL, Abcam Cambridge, cat no: ab12073), CDK2 (1:1000; Proteintech, USA, cat. no. 10122- 1- AP), CDK3 (1:2000; Proteintech, USA, cat. no. 55103- 1- AP), CDK6 (1:2000; Proteintech, USA, cat. no. 14052- 1- AP), cyclin D1 (1:5000; Proteintech, USA, cat. no. 26939- 1- AP), cyclin E1 (1:1000; Proteintech, USA, cat. no. 11554- 1- AP), p18 (1:1000, BOSTER China, cat. no. M03299- 1), p19 (1:1000, BOSTER China, cat. no. MA1075), p53 (1:5000; Proteintech, USA, cat. no. 60283- 2- Ig), p- p53 (1:2000; Proteintech, USA, cat. no. 28961- 1- AP), p57 (1:1000, BOSTER China, cat. no. BM4129), Rb (1:1000, BOSTER China, cat. no. BM4500), pRb (1:1000, BOSTER China, cat. no. BM4338), Bcl- 2 (1:1000; Proteintech, USA, cat. no. 26593- 1- AP), Bax (1:2000; Proteintech, USA, cat. no. 50599- 2- lg), Ecadherin (1:5000; Proteintech, USA, cat. no. 20874- 1- AP), N- cadherin (1:3000; Proteintech, USA, cat. no. 22018- 1- AP), vimentin (1:4000; Proteintech, USA, cat. no. 10366- 1- AP), MMP1 (1:1000, BOSTER China, cat. no. A00733- 1), MMP2 (1:500, BOSTER China, cat. no. BM4075), MMP9 (1:1000, BOSTER China, cat. no. PB0709), MMP14 (1:1000, BOSTER China, cat. no. BM4119), TIMP1 (1:1000, Bioss China, cat. no. bs0415R), TIMP2 (1:1000, Bioss China, cat. no. bs- 10395R), TIMP3 (1:1000; Proteintech, USA, cat. no. 10858- 1- AP), ITGB1 (1:1000, BOSTER China, cat. no. BM4308), ITGB3 (1:1000, BOSTER China, cat. no. BA1670), ITGB5 (1:1000, BOSTER China, cat. no. A04201- 1), nm23 (1:1000, BOSTER China, cat. no. BA3787), PD- L1 (1:3000; Proteintech, USA, cat. no. 66248- 1- Ig), and βactin (1:5000; Proteintech, USA, cat. no. 66009- 1- Ig).

Techniques: Expressing, Quantitative RT-PCR, Western Blot