Journal: BMC cancer

Article Title: Identification of the angiogenic gene signature induced by EGF and hypoxia in colorectal cancer.

doi: 10.1186/1471-2407-13-518

Figure Lengend Snippet: Figure 5 EGF receptor is autophosphorylated in Caco-2 and activates downstream signalling pathways. Caco-2 cells were stimulated with 20 ng/mL EGF for the time periods indicated. Western blotting for (a) phosphorylated EGFR or total EGFR and (b) antibodies recognising signal- ling enzymes is shown. Cell lysate of EGF-treated A431 cells was used as positive control. α-tubulin is shown as a loading control. Densitometry was performed using Phoretix 1D analysis software against α-tubulin (for ERK, data for p42 and p44 are shown).

Article Snippet: Whole cell lysate of EGF-treated A431 epithelial carcinoma cells used as positive control was from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Western Blot, Positive Control, Control, Software

Journal:

Article Title: Infection of Epithelial Cells by Pathogenic Neisseriae Reduces the Levels of Multiple Lysosomal Constituents

doi:

Figure Lengend Snippet: LAMP2 and EGFR levels in neisseria-infected A431 human epithelial cells. (A) LAMP2 and β-tubulin signals in representative immunoblots of total cell proteins from uninfected cultures and cultures infected with piliated bacteria. U, uninfected cultures; GC, WT piliated GC (strain GCM740); GC iga, GCM740Δ4, a protease-deficient but otherwise isogenic derivative of GCM740; MC, WT piliated MC (strain 8013.6). (B) LAMP2 levels in infected A431 cultures relative to that in the uninfected control culture (means and standard deviations [vertical bar] from four independent experiments). (A′) LAMP2 and β-tubulin signals in representative immunoblots of total cell proteins from A431 cells infected with MS11A307, a nonpiliated, Opa−, low-adherence strain. (B′) LAMP2 levels in A431 cultures infected with piliated MS11A (GC P+) and nonpiliated MS11A307 (GC P−) relative to the uninfected control culture. (C) EGFR level in A431 cultures infected with strain GCM740 (GC) expressed relative to that in the uninfected (control) culture (U) (mean and standard deviation [vertical bar] from three independent experiments).

Article Snippet: A431 human epidermoid carcinoma epithelial cells (ATCC CRL 1555) were maintained in Dulbecco’s minimum essential medium (Gibco) plus 5% fetal bovine serum (Gibco), and infections were carried out in the same medium plus 50 μg of human transferrin per ml.

Techniques: Infection, Western Blot, Bacteria, Control, Standard Deviation

Journal:

Article Title: Infection of Epithelial Cells by Pathogenic Neisseriae Reduces the Levels of Multiple Lysosomal Constituents

doi:

Figure Lengend Snippet: Confocal laser scanning microscopy of GC-infected A431 cells double-stained for LAMP2 (green) and bacteria (red). Panels A and B show one horizontal 4-μm-thick optical section taken of the same field of cells. The arrow indicates the location of a cell demonstrated by phase-contrast microscopy (data not shown) and by its fluorescence properties to be infected.

Article Snippet: A431 human epidermoid carcinoma epithelial cells (ATCC CRL 1555) were maintained in Dulbecco’s minimum essential medium (Gibco) plus 5% fetal bovine serum (Gibco), and infections were carried out in the same medium plus 50 μg of human transferrin per ml.

Techniques: Confocal Laser Scanning Microscopy, Infection, Staining, Bacteria, Microscopy, Fluorescence

Journal:

Article Title: Infection of Epithelial Cells by Pathogenic Neisseriae Reduces the Levels of Multiple Lysosomal Constituents

doi:

Figure Lengend Snippet: Biosynthesis rates of LAMP2 in uninfected A431 cell cultures (U) and cultures infected with WT GC (strain GCM740) (I). (A) A representative autoradiogram of LAMP2 immunoprecipitated from radiolabelled cultures. (B) Rates of incorporation of [35S]Met-[35S]Cys into LAMP2. Data are from three independent experiments.

Article Snippet: A431 human epidermoid carcinoma epithelial cells (ATCC CRL 1555) were maintained in Dulbecco’s minimum essential medium (Gibco) plus 5% fetal bovine serum (Gibco), and infections were carried out in the same medium plus 50 μg of human transferrin per ml.

Techniques: Infection, Immunoprecipitation

Journal:

Article Title: Infection of Epithelial Cells by Pathogenic Neisseriae Reduces the Levels of Multiple Lysosomal Constituents

doi:

Figure Lengend Snippet: LAP activities in cultures of A431 cells infected with MC strain 8013.6 (MC) or GC strains GCM740 (GC) and GCM740Δ4 (GC iga) expressed relative to that in control uninfected cells (U). Results represent the means and standard deviations from three independent experiments.

Article Snippet: A431 human epidermoid carcinoma epithelial cells (ATCC CRL 1555) were maintained in Dulbecco’s minimum essential medium (Gibco) plus 5% fetal bovine serum (Gibco), and infections were carried out in the same medium plus 50 μg of human transferrin per ml.

Techniques: Infection, Control

Journal:

Article Title: Infection of Epithelial Cells by Pathogenic Neisseriae Reduces the Levels of Multiple Lysosomal Constituents

doi:

Figure Lengend Snippet: CD63 levels in uninfected A431 cells (A and B) and cells infected with N. meningitidis 8013.6 (C and D), visualized by double-label immunofluorescence microscopy. (A and C) Cells stained for CD63. (B and D) The same field stained for MC. Stained cells were examined and photographed with a Nikon Microphot FX at the same magnifications. Note that the uninfected cell within the center of this field of infected cells (panel D) had normal levels of CD63 (panel C).

Article Snippet: A431 human epidermoid carcinoma epithelial cells (ATCC CRL 1555) were maintained in Dulbecco’s minimum essential medium (Gibco) plus 5% fetal bovine serum (Gibco), and infections were carried out in the same medium plus 50 μg of human transferrin per ml.

Techniques: Infection, Immunofluorescence, Microscopy, Staining

Journal: British Journal of Cancer

Article Title: Substituted titanocenes induce caspase-dependent apoptosis in human epidermoid carcinoma cells in vitro and exhibit antitumour activity in vivo

doi: 10.1038/sj.bjc.6604021

Figure Lengend Snippet: Antiproliferative effect of Titanocenes X and Y, Cp 2 TiCl 2 and cisplatin on HeLa and A431 cells. HeLa ( A and B ) and A431 ( C and D ) cells were treated with vehicle (DMSO (0.5% v/v)) or 0.5, 5, 50 and 250 μ M of the indicated compound for 24 h ( A and C ) and 48 h ( B and D ). Cell viability was determined using an MTT assay and absorbance was monitored spectrophotometrically at 540 nm. Results represent the mean±s.e.m. of three separate experiments. ( E ) IC50 values±s.e. from Cp 2 TiCl 2 , cisplatin, Titanocenes X and Y treatment of HeLa and A431 cells for 48 h, calculated using the PRISM statistical analysis software package.

Article Snippet: The HeLa cervical carcinoma cell line and the A431 carcinoma cell line were purchased from the American Tissue Culture Collection, LGC Promochem, Middlesex, UK.

Techniques: MTT Assay, Software

Journal: British Journal of Cancer

Article Title: Substituted titanocenes induce caspase-dependent apoptosis in human epidermoid carcinoma cells in vitro and exhibit antitumour activity in vivo

doi: 10.1038/sj.bjc.6604021

Figure Lengend Snippet: Effect of Titanocene Y and cisplatin on growth of A431 xenograft mice in vivo. A431 cells (1 × 10 7 ) were injected subcutaneously (s.c.) at day 0 into female NMRI:nu/nu mice (eight per group) allowed to reach 5–6 mm in diameter. Mice were treated i.p. with cisplatin (3 mg kg day −1 ) or Titanocene Y (40 mg kg day −1 ) for 5 consecutive days starting on day 8. For each treatment group, tumour volume was measured using calipers on days 10, 14, 17 and 21 ( A ) and mean body weight was calculated ( B ).

Article Snippet: The HeLa cervical carcinoma cell line and the A431 carcinoma cell line were purchased from the American Tissue Culture Collection, LGC Promochem, Middlesex, UK.

Techniques: In Vivo, Injection

Journal: British Journal of Cancer

Article Title: Substituted titanocenes induce caspase-dependent apoptosis in human epidermoid carcinoma cells in vitro and exhibit antitumour activity in vivo

doi: 10.1038/sj.bjc.6604021

Figure Lengend Snippet: A431 cells undergo apoptosis when treated with Titanocene Y. Cells (0.8 × 10 6 ) were treated with ( A ) vehicle (DMSO (0.5% v/v)) or ( B ) Titanocene Y (50 μ M ) or ( C ) cisplatin (50 μ M ) for 48 h. Cells were harvested by trypsination and centrifugation and stained with Annexin V/FITC antibody for 15 min according to the manufacturer's instructions. Cells were washed in binding buffer and stained with propidium iodide. Annexin V and propidium iodide fluorescence was measured by flow cytometry.

Article Snippet: The HeLa cervical carcinoma cell line and the A431 carcinoma cell line were purchased from the American Tissue Culture Collection, LGC Promochem, Middlesex, UK.

Techniques: Centrifugation, Staining, Binding Assay, Fluorescence, Flow Cytometry

Journal: British Journal of Cancer

Article Title: Substituted titanocenes induce caspase-dependent apoptosis in human epidermoid carcinoma cells in vitro and exhibit antitumour activity in vivo

doi: 10.1038/sj.bjc.6604021

Figure Lengend Snippet: Effect of Cp 2 TiCl 2 , cisplatin, Titanocene X and Titanocene Y on A431 and HeLa cell cycle. A431 ( A and B ) and HeLa ( C and D ) cells were treated with vehicle (DMSO) (0.5% v/v) or 50 μ M Cp 2 TiCl 2 , cisplatin, Titanocene X or Titanocene Y for ( A and C ) 24 h and ( B and C ) 48 h. DNA was stained with propidium iodide, and DNA content was analysed by flow cytometry. Results represent the mean±s.e.m. of three separate experiments.

Article Snippet: The HeLa cervical carcinoma cell line and the A431 carcinoma cell line were purchased from the American Tissue Culture Collection, LGC Promochem, Middlesex, UK.

Techniques: Staining, Flow Cytometry

Journal: British Journal of Cancer

Article Title: Substituted titanocenes induce caspase-dependent apoptosis in human epidermoid carcinoma cells in vitro and exhibit antitumour activity in vivo

doi: 10.1038/sj.bjc.6604021

Figure Lengend Snippet: Western blot analysis of caspase-3, -7 and -8 following Titanocene Y treatment of A431 cells. Cells were treated with either vehicle (0.5% DMSO (v/v)) or Titanocene Y (50 μ M ) for ( A ) 24 or 48 h or ( B ) a range of Titanocene Y concentrations (0–50 μ M ) for 48 h. Whole cell extracts were prepared and protein (60 μ g) and resolved by SDS–PAGE followed by Western blotting using antibodies to detect the active cleavage product of caspase-3, -7 and -8. As a positive control (+VE) for caspase 8 activity, PWR-1 prostate cells were treated with etoposide (10 μ M ) for 48 h. Blots were stripped and re-probed with β -actin or GAPDH as loading controls. Results are representative of three independent experiments. ( C ) A431 cells (1 × 10 5 cells) were treated with vehicle (DMSO (0.5% v/v)), 50 μ M Titanocene Y or 50 μ M cisplatin for 48 h, or a pretreatment of z-VAD-fmk (150 μ M ) for 1 h prior to DMSO (0.5% v/v) or 50 μ M Titanocene Y or cisplatin for a further 48 h. Cell viability was determined by flow cytometry following propidium iodide staining. The appearance of a pre-G 1 peak represents non-viable cells. * P <0.005 with respect to cisplatin treatment, Student's t -test. Results represent the mean±s.e.m. of three separate experiments.

Article Snippet: The HeLa cervical carcinoma cell line and the A431 carcinoma cell line were purchased from the American Tissue Culture Collection, LGC Promochem, Middlesex, UK.

Techniques: Western Blot, SDS Page, Positive Control, Activity Assay, Flow Cytometry, Staining