Journal: Kidney international

Article Title: R-roscovitine (CYC202) alleviates renal cell proliferation in nephritis without aggravating podocyte injury.

doi: 10.1111/j.1523-1755.2005.00213.x

Figure Lengend Snippet: Fig. 5. Immunostaining for cyclin D1 in rats with passive Heymann’s nephritis receiving either vehicle or CYC202 from day 3 to 30 after disease induction. (A) In vehicle-treated nephritic rats on day 9, cyclin D1 is expressed mostly in the cytoplasm (magnification ×400). (B) In nephritic rats receiving high-dose CYC202, the expression of cyclin D1 on day 9 is qualitatively similar as in rats receiving vehicle, although quantitatively reduced (magnification ×400). (C) Quantitative assess- ment of glomerular cells staining positive for cyclin D1. ∗P < 0.05 versus nephritic rats receiving vehicle alone. Rats receiving vehicle only (□) (N = 9); rats receiving 25 mg/kg/day of CYC202 ( ) (N = 9); rats receiving 50 mg/kg/day (■) of CYC202 (N = 8).

Article Snippet: Primary antibodies included [14]: (1) a monoclonal IgG1 antibody (clone D33) against human muscle desmin (1:100) (Dako, Glostrup, Denmark); (2) cyclin D1, a rabbit polyclonal antibody (bcl-1 Ab-3) (1:100) (Lab Vision, Fremont, CA, USA) raised against the C-terminal portion of cyclin D1 of rat origin [15]; (3) ED1 (1:500)(Serotec, Oxford, UK), a monoclonal IgG antibody to a cytoplasmic antigen present in monocytes, macrophages and dendritic rat cells; (4) a monoclonal IgG antibody (clone 1A4) to an NH2-terminal synthetic decapeptide of human alpha-smooth muscle actin (a-SMA) (1:500) (Dako); (5) BU-1, a monoclonal antibody against bromo-deoxyuridine (Amersham, Braunschweig, Germany); (6) proliferating cell nuclear antigen (PCNA) (Ab-1) (clone PC10) (1:1000) (Oncogene Science Inc., Uniondale, NY, USA) a mouse monoclonal IgG antibody that reacts with the human PCNA, which is expressed in a cell cycle dependent manner [16]; and (7) WT1 (F-6) (1:300) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), a mouse monoclonal IgG1 that reacts with the WT1 antigen on podocytes and epithelial cells of the Bowman capsule.

Techniques: Immunostaining, Expressing, Staining

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