Journal: World Journal of Surgical Oncology

Article Title: eIF6: a promising therapeutic target for gastric carcinoma via PI3K/AKT pathway modulation

doi: 10.1186/s12957-025-03746-w

Figure Lengend Snippet: Antibody

Article Snippet: CDK2 , Boster , PB9534 , 30 kDa , Rabbit.

Techniques:

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: Journal of molecular medicine (Berlin, Germany)

Article Title: Epigenetic silencing of PRSS3 provides growth and metastasis advantage for human hepatocellular carcinoma

doi: 10.1007/s00109-017-1578-5

Figure Lengend Snippet: Effect of PRSS3 on the growth properties of human HCC cells. HCC HepG2 and PLC/PRF/5 cell lines with overexpression of PRSS3, and SNU-387 cell line with knockdown of PRSS3 expression were established to evaluate the potential roles of PRSS3 in HCC development. a, b MTT assays showed the viability of HCC HepG2 and PLC/PRF/5 cells with PRSS3 overexpression (PRSS3) or vector control (Vector) (a), or SNU-387 cells transfected with either siRNA against PRSS3 (siPRSS3–2) [20] or RNAi Negative Control Duplex (siNC) (b). c, d. Colony formation in soft agar for 2 weeks by HCC HepG2 and PLC/PRF/5 cells with PRSS3 overexpression, or control (C), or SNU-387 cells transfected with siPRSS3–2 or siNC (d). Left panel: representative image; Right panel: quantitative analysis. e, f FACS analysis of cell cycle distribution of HepG2 and PLC/PRF/5 cells with or without ectopic expression of PRSS3 (E), or SNU-387 cells transfected with siPRSS3–2 or siNC (f). Left panel: representative FACS histograms with the percentage of cells in each cell cycle phase (Left red peak: G0/G1; right red peak: G2/M; hatched peak: S; coefficient of variation of G1 peak: % CV); right panel: quantitative graphs. g Western blotting analysis of the levels of PRSS3, cyclin D1, CDK4, cyclin E1, and CDK2 in the transfected HCC cell lines. The experiments were repeated at least three times, and the results were presented as the mean ± SD, *p < 0.05, versus control

Article Snippet: The cells were split to low density (30% confluence) for overnight culture and were then treated with 2 μM of 5-AZA (Sigma-Aldrich) for 96 h with the medium exchanged every 24 h or with 4 μM of trichostatin A (TSA) (Sigma-Aldrich) for 24 h. For combined treatment, the cells were initially exposed to 5-AZA for 72 h followed by 5-AZA and TSA for 24 h. The primary antibodies were used against the following proteins for Western blot: PRSS3 from R&D Systems (Cat. no.: MAB3710); p-MEK1/2 from Cell Signaling Technology (Cat. no.: 9121); cyclin D1 from Proteintech Group, Inc. (Cat. no.: 60186–1-Ig); and cyclin-dependent kinase 2 (CDK2), CDK4, cyclin E1, matrix metallopeptidase 2 (MMP2), MEK1/2, ERK1/2, p-ERK1/2, and β-actin from Bioworld Technology, Inc. (Cat. nos.

Techniques: Over Expression, Expressing, Plasmid Preparation, Transfection, Negative Control, Western Blot