Journal: Journal of ethnopharmacology

Article Title: Dehydrocorydaline attenuates myocardial ischemia-reperfusion injury via the FoXO signalling pathway: A multimodal study based on network pharmacology, molecular docking, and experimental study.

doi: 10.1016/j.jep.2024.118738

Figure Lengend Snippet: Fig. 5. Prediction and validation of DHC and its potential targets by molecular docking and IHC analysis. (A) Molecular docking of STAT3 and DHC. (B) Molecular docking of MDM2 and DHC. (C) Molecular docking of CDK2 and DHC. (D) Molecular docking of PLK1 and DHC. (E) Molecular docking of CCND1 and DHC. (F) Binding of DHC to CCND1 as determined through microscale thermophoresis (MST). (G) Binding of DHC to MDM2 as determined through MST. (H) Binding of DHC to CDK2 as determined through MST. (I) Relative protein levels of p-FOXO1A (n = 3). (J) Representative IHC images of p-FOXO1A, CCND1, and p-MDM2. (K) Relative protein levels of CCND1 (n = 3). (L) Relative protein levels of p-MDM2 (n = 3). *indicates a significant difference compared with the sham group, # indicates a significant difference compared with the MIRI group: *p < 0.05, **p < 0.01, #p < 0.05, ##p < 0.01. DHC: dehydrocorydaline, IHC: immunohistochemical, MIRI: myocardial ischemia-reperfusion injury.

Article Snippet: The microscale thermophoresis (MST) technique was performed to validate the interaction between CCND1 (#HY-P72119, MedChemExpress, Shanghai, China), CDK2 (#HY-P70014, MedChemExpress, Shanghai, China), MDM2 (#HY-P701593, MedChemExpress, Shanghai, China) and DHC(#HY-N0674, MedChemExpress, Shanghai, China).

Techniques: Biomarker Discovery, Binding Assay, Microscale Thermophoresis, Immunohistochemical staining

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

Journal: bioRxiv

Article Title: A cyclin-dependent kinase 5-derived peptide inhibits Cdk5/p25 activity and improves neurodegenerative phenotypes

doi: 10.1101/2020.05.12.090472

Figure Lengend Snippet: A , Strategy for the design of Cdk5/p25 complex-specific inhibitory peptide (Cdk5i peptide). The structure of Cdk5/p25 complex was downloaded from RCSB protein data bank. B, Microscale thermophoresis-based biophysical assays assessing binding of Cdk5i peptide to recombinant proteins. C , Recombinant Cdk5/p25 complex was incubated with biotin-conjugated Cdk5i or scrambled peptide for overnight at 4°C, and subjected to pull-down assay with streptavidin-coated beads. Interaction between the peptides and Cdk5, or p25 was addressed by immunoblotting. D, Brain lysates from wild-type mice were incubated with Cdk5i peptide or scrambled for overnight at 4°C, and subjected to pull-down assay. Interaction between the peptides and Cdk5, Cdk2 or p35 was addressed by immunoblotting.

Article Snippet: 6X-HIS tagged CDK5 (CDK5-236H), and 6X-HIS tagged Cdk2 (CDK2-655H) were purchased from Creative Biomart ( https://www.creativebiomart.net/ ).

Techniques: Microscale Thermophoresis, Binding Assay, Recombinant, Incubation, Pull Down Assay, Western Blot

Journal: bioRxiv

Article Title: A cyclin-dependent kinase 5-derived peptide inhibits Cdk5/p25 activity and improves neurodegenerative phenotypes

doi: 10.1101/2020.05.12.090472

Figure Lengend Snippet: A, Recombinant Cdk5/p25 complex was incubated with Cdk5i peptide or scrambled overnight at 4°C, and subjected to IP-linked Cdk5 kinase assay. The bar graph represents the relative radioactivity of 32 P incorporated into histone H1. B, Brain lysates from three-month-old Tau P301S mice were incubated with Cdk5i peptide or scrambled overnight at 4°C, and subjected to IP-linked Cdk5 kinase assay. C-D, Brain lysates from three-month-old wild-type mice were incubated with Cdk5i peptide or scrambled overnight at 4°C, and subjected to IP-linked kinase assay for Cdk5 or Cdk2.

Article Snippet: 6X-HIS tagged CDK5 (CDK5-236H), and 6X-HIS tagged Cdk2 (CDK2-655H) were purchased from Creative Biomart ( https://www.creativebiomart.net/ ).

Techniques: Recombinant, Incubation, Kinase Assay, Radioactivity

Journal: Poultry Science

Article Title: Dietary naringin supplementation on laying performance and antioxidant capacity of Three-Yellow breeder hens during the late laying period

doi: 10.1016/j.psj.2022.102023

Figure Lengend Snippet: Ovarian morphology, index, and protein expression of Three-Yellow breeder hens . a,b Mean values with different superscripts letters denote statistical significantly difference ( P < 0.05). (A) Ovary morphology in each group; (B) Numbers of the preovulatory follicles (with diameters over 12 mm) in each group (n = 12). (C) Ovarian histomorphology in each group, scale bar: 50 μm. (D) Ovarian index in each group (n = 12). (E) Western blot analysis of CDK2 and PCNA in ovary. F: Expression levels of proteins related to cell proliferation of ovarian tissues in each group. Abbreviations: C, basal diet group; N1, N2, and N3, basal diet supplemented with 0.1%, 0.2%, and 0.4% of naringin, respectively.

Article Snippet: After blocked with 5% skimmed milk at room temperature for 2 h, the blots were probed with corresponding primary antibodies β-actin (1:1000; BM3873), PCNA (1:1000; BM0104) and CDK2 (1:1000; PB0562) purchased from Boster Biological Technology Co. Ltd., (Wuhan, China) with optimized conditions, and then incubated with the secondary antibody.

Techniques: Expressing, Western Blot