Journal: Carcinogenesis

Article Title: Wntless (GPR177) expression correlates with poor prognosis in B-cell precursor acute lymphoblastic leukemia via Wnt signaling.

doi: 10.1093/carcin/bgu166

Figure Lengend Snippet: Fig. 4. Wls expression is critical for Wnt-induced consequent downstream genes’ activation. (A) Ablated Wls protein expression in B-cell leukemia cell lines transfected with Wls shRNAi. (B) Decrease of Wls expression in Reh cells infected with Wls shRNAi was showed to abrogate canonical Wnt signaling induced by Wnt-3a stimulation. (C) Inhibition of c-Myc, cyclin D1 and Skp2 expression induced by Wnt-1 and Wnt-3a in the Reh cells infected with Wntless shRNAi. Arrow head, cyclin D1. (D) Reh cells infected with Wntless shRNAi showed a decrease in pro-inflammatory cytokines (IL-1β, IL-6, IL-8, GM-CSF and cox-2) expression induced by Wnt-3a treatment. Each experiment was repeated at least three. The results are shown as means ± SD. (***p < 0.001).

Article Snippet: The primary antibodies used in this study were β-actin polyclonal antibody (1:2000, Santa Cruz, I-19), cIAP, Bcl-xL polyclonal antibody (1:2000, Santa Cruz), caspase 3, caspase 8, c-Myc (1:1000, Upstate), cyclin D1 (SeroTec.), FITC-conjugated anti-mouse, alkaline phosphatase-conjugated anti-rabbit antibodies (1:500, Jackson ImmunoResearch Lab.) and Wntless antibody (1:2000, Biolegend) (22).

Techniques: Expressing, Activation Assay, Transfection, Infection, Inhibition

Journal: Cancer Science

Article Title: Novel MEK inhibitor trametinib and other retinoblastoma gene (RB)‐reactivating agents enhance efficacy of 5‐fluorouracil on human colon cancer cells

doi: 10.1111/cas.12139

Figure Lengend Snippet: Fenofibrate induces G1‐phase arrest through a blockade of the mitogen‐activated protein kinase (MAPK) cascade in HT‐29 cells. (a) Growth inhibitory effect of fenofibrate on HT‐29 cells. Cells were treated with fenofibrate at the indicated concentrations for 72 h, and viability was measured with a Cell Counting Kit‐8 assay. The data obtained with the solvent dimethyl sulfoxide (DMSO) was taken as 100%. Points, means (n = 3); bars, standard deviation (SD). **P < 0.01, compared with the DMSO‐treated control. (b) Cell cycle analysis of HT‐29 cells treated with fenofibrate. Cells were treated with fenofibrate at the indicated concentrations for 24 h. The DNA contents of the cells were analyzed by flow cytometry. The percentage in each phase of the cell cycle is shown. Columns, means (n = 3); bars, SD. **P < 0.01, compared with the DMSO‐treated control. (c) The effect of fenofibrate at 100 μΜ for 24 h on the phosphorylation status of ERK and the expression of cyclin D1 analyzed by Western blotting. α‐tubulin was used as a loading control.

Article Snippet: The following were used as the primary antibody: rabbit anti‐human p15 polyclonal antibody (C‐20, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti‐human p21 polyclonal antibody (C‐19, Santa Cruz Biotechnology), rabbit anti‐human p27 polyclonal antibody (C‐19, Santa Cruz Biotechnology), mouse anti‐human cyclin D1 monoclonal antibody (MBL, Nagoya, Japan), mouse anti‐human thymidylate synthase monoclonal antibody (Abcam, Cambridge, UK), rabbit anti‐human Akt, phospho‐Akt (Ser473), p42/44 MAPK, phospho‐p42/44 MAPK, phospho‐Rb (Ser780), and phospho‐Rb (Ser807/811) (Cell Signaling Technology).

Techniques: Cell Counting, Standard Deviation, Cell Cycle Assay, Flow Cytometry, Expressing, Western Blot

Journal: PLoS ONE

Article Title: Grb7 Protein Stability Modulated by Pin1 in Association with Cell Cycle Progression

doi: 10.1371/journal.pone.0163617

Figure Lengend Snippet: (A) Cell lysates from shLuc- or shPin1-infected (shPin1 #1 or shPin1 #2) A431 cells were collected and subjected to Western blotting with anti-Pin1, anti-Grb7, or anti-Cyclin D1 antibody to examine the protein expression of the indicated molecules. (B) Cell lysates from Pin1 wild-type (Pin1 +/+ ) and knockout (Pin1 -/- ) mouse embryonic fibroblasts were collected and subjected to Western blotting with anti-Pin1 or anti-Grb7 antibody. (C) Cell lysates from shLuc-, shPin1-, or shGrb7-infected A431 cells were collected and subjected to Western blotting with anti-Pin1 or anti-Grb7 antibody. The results demonstrated that the protein expression of Grb7 was increased in the Pin1 knockdown condition. Each experiment was repeated at least three independent times and the representative blots were shown in A-C.

Article Snippet: The mouse monoclonal anti-Cyclin D1 (5D4, ADI-KAM-CC200-E) antibody was purchased from Enzo Life Science (Farmingdale, NY).

Techniques: Infection, Western Blot, Expressing, Knock-Out