Journal: Experimental Biology and Medicine

Article Title: Liraglutide prevents fast weight gain and β-cell dysfunction in male catch-up growth rats

doi: 10.1177/1535370214567614

Figure Lengend Snippet: Effects of HFD catch-up growth and liraglutide treatment on β-cell function and apoptosis. Pdx-1 protein (a1) and mRNA (a2) levels of islets at the end of week 8. Pdx-1 protein levels of islets were assessed by Western blot and the results were expressed as the ratios of Pdx-1 and GAPDH. The Pdx-1 mRNA levels were detected by qPCR and expressed as the ratios of Pdx-1 and β-actin. Protein levels of Bcl-2 (b1) and Caspase-3 p 12 subunit (c1) were detected by Western blot and the results were expressed as the ratios of Caspase-3 p12 subunit or Bcl-2 with GAPDH. The mRNA levels of Bcl-2 (b2) and Procaspase-3 (c2) were expressed as the ratios of Proaspase-3 or Bcl-2 with β-actin. All the results are expressed as mean ± SEM. *p < 0.01 versus the NC group; #p < 0.01 versus the CUG group; &p < 0.01 versus the CUGL group

Article Snippet: Primer sequences Western blot analysis Protein extracts from islets (20 μg from each rat) were subjected to Western blot analysis using mouse antipancreatic duodenal homeobox-1 (Pdx-1) monoclonal antibody (1:800; Cell Signaling Technology, Danvers, MA, USA), mouse anti-Caspase-3 p12 subunit monoclonal antibody (1:500, Boster Bio-Engineering, China), mouse anti-B-cell lymphoma (Bcl-2) polyclonal antibody (1:800, Bioworld, USA), and mouse anti-GAPDH antibody (1:1000, Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Cell Function Assay, Western Blot

Journal: Molecular Cancer

Article Title: Arecoline induces HA22T/VGH hepatoma cells to undergo anoikis - involvement of STAT3 and RhoA activation

doi: 10.1186/1476-4598-9-126

Figure Lengend Snippet: Arecoline alters the expression of apoptosis-related proteins and caspase activity in HA22T/VGH cells . (A) HA22T/VGH cells were treated with 0, 30, or 100 μg/ml of arecoline for 24 h, then the cells were harvested and proteins extracted and used for Western blotting for Bcl-2, Bcl-X L , Bax, cytochrome c, or procaspase-9. β-actin was used as the internal control. The values shown are the quantitative density analysis expressed as the relative density compared to that in untreated cells (control), taken as 100%. The results are expressed as the mean ± S.D. for three separate experiments. (B) Caspase-3 activity was detected using RPE-conjugated anti-active caspase-3 antibody by flow cytometric analysis. The values shown are the percentage of cells with active caspase-3 and are the mean ± S.D. of three independent experiments. The red filled area is the untreated control and the black lines the treated groups. *: p < 0.05 as compared to the untreated control.

Article Snippet: R-phycoerythrin (RPE)-conjugated rabbit anti-active caspase-3 polyclonal antibodies, RPE-conjugated mouse anti-human β1-integrin monoclonal antibody and the RPE-conjugated mouse IgG isotype control were purchased from BD Pharmingen Inc. (San Diego, CA, USA).

Techniques: Expressing, Activity Assay, Western Blot

Journal: Molecular Cancer

Article Title: Arecoline induces HA22T/VGH hepatoma cells to undergo anoikis - involvement of STAT3 and RhoA activation

doi: 10.1186/1476-4598-9-126

Figure Lengend Snippet: Schematic representation of the arecoline-stimulated signaling pathways for detachment and apoptosis of HA22T/VGH cells . Arecoline treatment decreases IL-6 levels, but does not change gp130 of IL-6 receptor. In addition, phosphorylation/activation of STAT3, which provides protection against anoikis, is inhibited and levels of its downstream signals IL-6, Bcl-2, and Bcl-X L are decreased, while Bax levels, mitochondrial cytochrome c release, caspase-9 levels, and caspase-3 activity are increased. Phosphorylation/activation of p190RhoGAP, a RhoA inhibitor, and its upstream regulator, SHP2, are inhibited, while the activation/cleavage of Rock-1, a RhoA downstream kinase, and actin stress fiber formation are increased, contributing to anoikis.

Article Snippet: R-phycoerythrin (RPE)-conjugated rabbit anti-active caspase-3 polyclonal antibodies, RPE-conjugated mouse anti-human β1-integrin monoclonal antibody and the RPE-conjugated mouse IgG isotype control were purchased from BD Pharmingen Inc. (San Diego, CA, USA).

Techniques: Activation Assay, Activity Assay

Journal: Pharmaceuticals

Article Title: Selenylated Imidazo [1,2- a ]pyridine Induces Apoptosis and Oxidative Stress in 2D and 3D Models of Colon Cancer Cells

doi: 10.3390/ph16060814

Figure Lengend Snippet: Caspase-3/7 activities in colon cancer cells treated with MRK-107. ( A ) Fluorescent signals of the negative control (NC) and Caco-2 and HT-29 cells at 24 h after treatment with MRK-107 or doxorubicin (DOX, positive control), as assessed using the CellEvent kit. ( B ) Caspase-3 activity in 3D cultures of Caco-2 and HT-29 at 24 h after treatment, as assessed by flow cytometry. Scale bar = 20 µm.

Article Snippet: For spheroids, labeling was performed using the PE Rabbit Anti-Active Caspase-3 kit (BD Pharmingen™, San Diego, CA, USA).

Techniques: Negative Control, Positive Control, Activity Assay, Flow Cytometry

Journal: Journal of Immunology (Baltimore, Md. : 1950)

Article Title: Mechanism for Pre-B Cell Loss in V H -Mutant Rabbits

doi: 10.4049/jimmunol.1101778

Figure Lengend Snippet: Flow cytometric analysis of ali/ali and wt B-lineage cell survival 2–6 d after transfer of pro-B cells to OP9 stroma in the absence of IL-7. A, Relative number of B-lineage (CD79a+) cells normalized to ali/ali cultures using bead-based counting and gating strategy in Fig. 2A. Error bars show SD from three independent experiments. B, Live/Dead staining of total lymphocytes. C, Cross-reactivity of anti–caspase-3 Ab with a UV-irradiated rabbit B cell line (55D1). Left panel, UV-treated 55D1 B cells immediately after irradiation (t0) (gray filled) or 3 h postirradiation stained with anti–caspase-3 (black dashed line) or isotype control for 3 h time point (solid black line) (identical result was observed for isotype control staining at t0); number is percentage of caspase-3+ cells at 3 h. Right panel, CD79a expression in caspase-3+ (dashed line) and caspase-3neg (solid line) cells 3 h postirradiation. D, Frequency of active caspase-3+ B-lineage cells (CD79a+). Data for B and D are representative of three independent experiments.

Article Snippet: Abs and flow cytometry Abs used: mouse anti-human CD79a-PE (catalog number 555935) or APC (catalog number 551134), rabbit anti-human caspase-3–FITC (catalog number 559341), mouse anti-human spleen tyrosine kinase (Syk)-P-Y348 (I1s0-722), mouse anti-rabbit–μ (367.2-bio), and rat anti-mouse pre-BCR (SL-156-bio) from BD Pharmingen (San Diego, CA); rat anti-mouse–μ-PE and PE-Cy7 (II/41) from eBioscience (San Diego, CA); goat F(ab′) 2 anti-human IgH+L, rabbit F(ab)-FITC anti-goat IgG H+L, and allophycocyanin-conjugated streptavidin from Jackson ImmunoResearch Laboratories (West Grove, PA); and biotinylated rabbit anti–a2-allotype and FITC-rabbit anti–a2-allotype IgG (generated by K.L.K.’s laboratory).

Techniques: Staining, Irradiation, Expressing