Journal: Anatomy & Cell Biology

Article Title: Expression and roles of NUPR1 in cholangiocarcinoma cells

doi: 10.5115/acb.2012.45.1.17

Figure Lengend Snippet: Nuclear protein-1 (NUPR1) affected epidermal growth factor (EGF)- or serum-induced invasion of HuCCT1 cholangiocarcinoma cells. To examine the effects on invasion, matrigel invasion assays were performed. Two days after siRNA transfection, 5×10 4 cells were seeded into matrigel-coated transwells. To rule out effects of proliferation, mitomycin C (5 µg/ml) was added. Eighteen hours later, the cells were fixed, stained with Diff-quik solution, and pictured. The representative staining of invaded cells is presented (A). Scale bar=50 µm. (B) Invaded cells were counted and the data are presented as a graph. Data show means±SDs of three independent experiments performed in triplicate ( * P <0.01). FBS, fetal bovine serum; SCR, scrambled.

Article Snippet: As described by Baek et al. [ ], following two days of transfection with SCR or NUPR1 siRNA into cells, transfected cells were seeded to a 24-well BioCoat Matrigel chamber inserts (BD Biosciences, San Jose, CA, USA).

Techniques: Transfection, Staining, Diff-Quik

Journal: Disease Models & Mechanisms

Article Title: Phenotype-driven chemical screening in zebrafish for compounds that inhibit collective cell migration identifies multiple pathways potentially involved in metastatic invasion

doi: 10.1242/dmm.018689

Figure Lengend Snippet: Confirmation of active compounds. (A) Schematic view of a 3 day post-fertilization (dpf) larva. The boxed area corresponds to the horizontal myoseptum (dotted red lines) and shows the area where leukocytes were counted in all quantification experiments. (B) Quantification of infiltrating leukocytes in the lateral line after treatments with diverse inhibitors. The assays were carried out by manually counting leukocytes recruited to the lateral line neuromasts after Cu 2+ treatment (10 μM) as described in Results. In this experiment, drugs were added 30 min prior to Cu 2+ treatment. The graph shows average leukocyte numbers in the lateral line in negative controls (DMSO), in Cu 2+ (as CuSO 4 ) and in Cu 2+ -plus drug-treated fish. Leukocyte quantification shows significant reduction (*** P <0.001) in neutrophil recruitment to damaged PLL neuromasts in larvae exposed to the compounds tested compared with Cu 2+ -exposed larvae, suggesting an inhibition of the neutrophil migration. Experiments were carried out with 15 larvae per condition. (C) Effects of kinase inhibitor compounds from LOPAC1280 on in vitro melanoma cell migration/invasion. 501-mel human melanoma cells were treated with DMSO (negative control) or the c-Met inhibitor K252a (positive control), and with the drugs indicated for 16 h and then assayed for motility as described in Materials and Methods. Representative data from independent experiments are shown. Only compounds that had a negative effect on migration are shown. (D) Quantification of 501-mel cell migration assays. The migratory cells were counted and the results expressed as the mean number of migratory cells from two independent experiments. Measurements were calculated relative to control (DMSO). (E) Comparative effects of tested drugs on the cell invasion capacity of melanoma cells. 501-mel cells were treated with DMSO, K252a, Wortmannin, Emodin and SU6656 and then plated on Matrigel inserts in presence of HGF at 50 ng/ml for 24 h. The invasive cells were counted and the results expressed as the mean number of migratory cells in five random microscopic fields. Measurements were calculated relative to control (DMSO). Data collected were from two independent experiments. *** P <0.001; *0.01< P <0.05.

Article Snippet: Invasion of cells into Matrigel was determined in vitro using 24-well BioCoat Matrigel inserts (Becton Dickinson) as described in ).

Techniques: Inhibition, Migration, In Vitro, Negative Control, Positive Control