Journal: Saudi journal of biological sciences

Article Title: Phytochemical profiling, antioxidant and antiproliferation potential of Euphorbia milii var.: Experimental analysis and in-silico validation.

doi: 10.1016/j.sjbs.2020.08.003

Figure Lengend Snippet: Fig. 2. Effect of E. milii on mRNA expression of specific gene of cell cycle regulators. (A) qRT-PCR was performed to assess the mRNA levels of cells regulators genes. HepG2 cells were cultured in the presence or absence of 11.2 mM E. milii water extract for 24 h. The GAPDH was used as internal positive control. The data for E. milii-treated samples were presented as the mean ± SEM of triplicate determination and compared with the control group. *p < 0.05; **p < 0.01. (B) Effect of E. milii on expression of cell cycle regulators. The cells were treated with or without E. milii for indicated time. The expression of CDK2, E2F1 and cyclin E were determined with western blot analysis.

Article Snippet: The nitrocellulose membranes were put into the blocking solution (5% fat free milk) for 1 h, then incubated the monoclonal anti-rabbit CDK2, cyclin E, E2F1 and anti-b-actin with shaking over night at 4 C. The horseradish peroxidase (HRP)conjugated secondary antibody given for at least 1 h, then chemiluminescence was detected using a chemiluminescence imaging system (Bio-Rad, Hercules, CA).

Techniques: Expressing, Quantitative RT-PCR, Cell Culture, Positive Control, Control, Western Blot

Journal: Saudi journal of biological sciences

Article Title: Phytochemical profiling, antioxidant and antiproliferation potential of Euphorbia milii var.: Experimental analysis and in-silico validation.

doi: 10.1016/j.sjbs.2020.08.003

Figure Lengend Snippet: Fig. 4. Obtained binding modes of ligands in the binding domain of CDK2 (magenta) and Thymidlate synthase (skyblue). (A) Binding mode of compound BAN in CDK2 ATP binding site. (B) Compound CBT bonded to CDK2. (C) Docking complex of CDK2-MBT. Binding mode of selected compounds in the active site of Thymidlate synthase (TS): (D) BAN-TS, (E) CBT-TS, (F) MBT-TS and (G) compound 5-FU (standard) bonded to TS.

Article Snippet: The nitrocellulose membranes were put into the blocking solution (5% fat free milk) for 1 h, then incubated the monoclonal anti-rabbit CDK2, cyclin E, E2F1 and anti-b-actin with shaking over night at 4 C. The horseradish peroxidase (HRP)conjugated secondary antibody given for at least 1 h, then chemiluminescence was detected using a chemiluminescence imaging system (Bio-Rad, Hercules, CA).

Techniques: Binding Assay

Journal: Saudi journal of biological sciences

Article Title: Phytochemical profiling, antioxidant and antiproliferation potential of Euphorbia milii var.: Experimental analysis and in-silico validation.

doi: 10.1016/j.sjbs.2020.08.003

Figure Lengend Snippet: Fig. 5. RMSDs of Ca atoms of the protein, backbone atoms of binding pocket (within 6.5 Å), and the heavy atoms in the ligand for: (A) CDK2-BAN, (B) CDK2-CBT, (C) TS-CBT and (D) TS-5-FU. Comparison between binding free energy terms of: (E) CDK2 bonded to BAN and CBT; (F) TS bonded to CBT and 5-FU.

Article Snippet: The nitrocellulose membranes were put into the blocking solution (5% fat free milk) for 1 h, then incubated the monoclonal anti-rabbit CDK2, cyclin E, E2F1 and anti-b-actin with shaking over night at 4 C. The horseradish peroxidase (HRP)conjugated secondary antibody given for at least 1 h, then chemiluminescence was detected using a chemiluminescence imaging system (Bio-Rad, Hercules, CA).

Techniques: Binding Assay, Comparison

Journal: The Journal of Biological Chemistry

Article Title: FAM129B/MINERVA, a Novel Adherens Junction-associated Protein, Suppresses Apoptosis in HeLa Cells *

doi: 10.1074/jbc.M110.175273

Figure Lengend Snippet: Intracellular localization of FAM129B in exponential and confluent cell cultures. A, HeLa cells (5 × 106) were harvested during the late exponential growth phase, and the cell extracts were fractionated into cytoplasmic (Cyt) and nuclear (Nuc) fractions (as described under “Experimental Procedures”). The fractions were analyzed by immunoblotting using antibodies directed against FAM129B, PARP, p53, CDK6, and CDK2. B, the same procedure was used to analyze NIH3T3 cell fractions. C, immunofluorescence microscopy of exponentially growing HeLa cells stained with a Hoechst 33342 DNA stain and with antibodies directed against rabbit FAM129B and the secondary antibody, chicken anti-rabbit IgG Alexa Fluor 594. The same protocol was followed for the cells in confluent HeLa cells (D) and an exponentially growing HeLa culture showing some mitotic cells (E). F, immunofluorescence co-localization (as described under “Experimental Procedures”) of FAM129B and β-catenin. The confluent cells were co-stained with rabbit antibodies directed against FAM129B and mouse antibodies directed against β-catenin. The secondary antibodies were Alexa Fluor 594-conjugated anti-rabbit IgG and an Alexa Fluor 488-conjugated anti-mouse IgG antibody. The cells were also stained with Hoechst 33342.

Article Snippet: Antibodies and Reagents Antibodies used for this study were rabbit anti-FAM129B (catalog no. 5122), rabbit anti-caspase-3 (catalog no. 9662), rabbit anti-cleaved caspase-3 (catalog no. 9664), rabbit anti-poly(ADP-ribose) polymerase (PARP 3 ; catalog no. 9542), rabbit anti-caspase-9 antibody (catalog no. 9502), mouse monoclonal anti-cdk6 (catalog no. 3136), mouse anti-caspase-8 (catalog no. 9746) (Cell Signaling, Beverly, MA); rabbit anti-cdk2 (catalog no. 21111) and rabbit anti-Akt (catalog no. 21054) (Signalway Antibody, Pearland, TX), mouse monoclonal antibody anti-β-catenin (catalog no. 610154) (BD Transduction Laboratories), mouse monoclonal anti-p53 (catalog no. sc-126), mouse monoclonal anti-GFP (catalog no. sc-9996), and mouse monoclonal β-tubulin (catalog no. sc-5274) (Santa Cruz Biotechnology, Inc., Santa Cruz).

Techniques: Western Blot, Immunofluorescence, Microscopy, Staining