Journal: International journal of cancer

Article Title: Specific induction of gadd45 in human melanocytes and melanoma cells after UVB irradiation.

doi: 10.1002/ijc.10275

Figure Lengend Snippet: FIGURE 1 – Kinetics of the p53 protein expression (detected by immunoblotting) after UVB 150J/m2 or UVC 50 J/m2 irradiation in p53 wildtype (A375P) and mutant p53 (SK-MEL2 and SK-MEL28) melanoma cells.

Article Snippet: Normal human melanocytes (NHM) were obtained and cultured as described previously.8 Human melanoma cell lines A375P, WM115, SK-Mel2, SK-Mel28, MeWo and Malme3M were obtained from the American Type Culture Collection.

Techniques: Expressing, Western Blot, Irradiation, Mutagenesis

Journal: International journal of cancer

Article Title: Specific induction of gadd45 in human melanocytes and melanoma cells after UVB irradiation.

doi: 10.1002/ijc.10275

Figure Lengend Snippet: FIGURE 2 – Effect of UVB (150 J/m2) or UVC (50 J m2) irradiation on the growth of A375P cells. Cells were plated in 24-well plate at a density of 2,500 cells/well. Alamar blue was added in medium 24, 48, 72 and 96 hr after beginning of treatment and fluorescence was read 4 hr later.

Article Snippet: Normal human melanocytes (NHM) were obtained and cultured as described previously.8 Human melanoma cell lines A375P, WM115, SK-Mel2, SK-Mel28, MeWo and Malme3M were obtained from the American Type Culture Collection.

Techniques: Irradiation

Journal: International journal of cancer

Article Title: Specific induction of gadd45 in human melanocytes and melanoma cells after UVB irradiation.

doi: 10.1002/ijc.10275

Figure Lengend Snippet: FIGURE 3 – Quantitative RT-PCR analysis of mdm2, p21WAF1, cyclin G1, gadd45, bax 16 hr after UVB 150 J/m2 and UVC 50 J/m2 irradiation in A375P melanoma cells. Results for all cells tested are summarized in Tables I and II.

Article Snippet: Normal human melanocytes (NHM) were obtained and cultured as described previously.8 Human melanoma cell lines A375P, WM115, SK-Mel2, SK-Mel28, MeWo and Malme3M were obtained from the American Type Culture Collection.

Techniques: Quantitative RT-PCR, Irradiation

Journal: International journal of cancer

Article Title: Specific induction of gadd45 in human melanocytes and melanoma cells after UVB irradiation.

doi: 10.1002/ijc.10275

Figure Lengend Snippet: FIGURE 4 – Increase of protein expression of gadd45 (detected by immunoblotting) in M4Be and A375P melanoma cells after UVB irradiation (150J/m2) or UVC irradiation (50J/m2) (NT untreated control).

Article Snippet: Normal human melanocytes (NHM) were obtained and cultured as described previously.8 Human melanoma cell lines A375P, WM115, SK-Mel2, SK-Mel28, MeWo and Malme3M were obtained from the American Type Culture Collection.

Techniques: Expressing, Western Blot, Irradiation, Control

Journal: International journal of cancer

Article Title: Specific induction of gadd45 in human melanocytes and melanoma cells after UVB irradiation.

doi: 10.1002/ijc.10275

Figure Lengend Snippet: FIGURE 5 – Quantitative RT-PCR analysis of mdm2, p21WAF1, cyclin G1, gadd45, bax 16 hr after UVB treatment with different doses in A375P melanoma cells. Results of quantitative analysis are in Table III.

Article Snippet: Normal human melanocytes (NHM) were obtained and cultured as described previously.8 Human melanoma cell lines A375P, WM115, SK-Mel2, SK-Mel28, MeWo and Malme3M were obtained from the American Type Culture Collection.

Techniques: Quantitative RT-PCR

Journal:

Article Title: Pharmacological and biochemical characterization of adenosine receptors in the human malignant melanoma A375 cell line

doi: 10.1038/sj.bjp.0704352

Figure Lengend Snippet: Expression of the adenosine receptors and β-actin transcripts in A375 cells. The PCR product in lane 2 is 394 bp long according to the human A1 receptor sequence, that in lane 3 is 208 bp long according to the human A2A receptor sequence, that in lane 4 is 153 bp long according to the human A2B receptor sequence and that in lane 5 is 156 bp long according to the human A3 receptor sequence. Lane 6 is 654 bp long according to the human β-actin sequence. Lanes 1 and 8 are molecular weight markers. Total RNA was extracted as described in Methods and used for RT – PCR (lanes 2 – 6) or directly used for PCR to detect β-actin DNA contaminant sequences (lane 7). Ten μl of PCR product were loaded in each lane. RT – PCR products obtained from A375 cells and separated on a 2% agarose gel.

Article Snippet: A375 cells (obtained from ATCC) were grown adherently and maintained in Dulbecco's Modified Eagle's Medium (DMEM), containing 10% foetal calf serum, penicillin (100 u ml −1 ), streptomycin (100 μg ml −1 ), L -glutamine (2 m M ) at 37°C in 5% CO 2 /95% air.

Techniques: Expressing, Sequencing, Molecular Weight, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis

Journal:

Article Title: Pharmacological and biochemical characterization of adenosine receptors in the human malignant melanoma A375 cell line

doi: 10.1038/sj.bjp.0704352

Figure Lengend Snippet: (A – C) Kinetics of [3H]-DPCPX, [3H]-SCH 58261 and [3H]-MRE 3008F20 binding to human A1, A2A and A3 adenosine receptors in A375 cells with association curves representative of a single experiment, which was replicated three times with similar results. The inset shows the data transformed as a semi-log plot, where Be is the specific binding measured at equilibrium and B is the specific binding at time t. (D – F) Kinetics of [3H]-DPCPX, [3H]-SCH 58261 and [3H]-MRE 3008F20 binding to human A1, A2A and A3 adenosine receptors in A375 cells with dissociation curves representative of a single experiment, which was replicated three times with similar results. The inset shows the data transformed as a semi-log plot, where B is the specific binding at time t and Be is the specific binding measured at equilibrium.

Article Snippet: A375 cells (obtained from ATCC) were grown adherently and maintained in Dulbecco's Modified Eagle's Medium (DMEM), containing 10% foetal calf serum, penicillin (100 u ml −1 ), streptomycin (100 μg ml −1 ), L -glutamine (2 m M ) at 37°C in 5% CO 2 /95% air.

Techniques: Binding Assay, Transformation Assay

Journal:

Article Title: Pharmacological and biochemical characterization of adenosine receptors in the human malignant melanoma A375 cell line

doi: 10.1038/sj.bjp.0704352

Figure Lengend Snippet: Saturation of [3H]-DPCPX, [3H]-SCH 58261 and [3H]-MRE 3008F20 binding to A1, A2A and A3 adenosine receptors in A375 cells. Experiments were performed as described in Methods. Values are the means and vertical lines s.e. of the mean of four separate experiments performed in triplicate. In the inset the Scatchard plot of the same data is shown.

Article Snippet: A375 cells (obtained from ATCC) were grown adherently and maintained in Dulbecco's Modified Eagle's Medium (DMEM), containing 10% foetal calf serum, penicillin (100 u ml −1 ), streptomycin (100 μg ml −1 ), L -glutamine (2 m M ) at 37°C in 5% CO 2 /95% air.

Techniques: Binding Assay

Journal:

Article Title: Pharmacological and biochemical characterization of adenosine receptors in the human malignant melanoma A375 cell line

doi: 10.1038/sj.bjp.0704352

Figure Lengend Snippet: Competition curves of specific [3H]-DPCPX, [3H]-SCH 58261 and [3H]-MRE 3008F20 binding to human A1, A2A and A3 adenosine receptors in A375 cells by adenosine agonists (A,C,E) and antagonists (B,D,F). Curves are representative of a single experiment from a series of four independent experiments. Competition experiments were performed as described in Methods.

Article Snippet: A375 cells (obtained from ATCC) were grown adherently and maintained in Dulbecco's Modified Eagle's Medium (DMEM), containing 10% foetal calf serum, penicillin (100 u ml −1 ), streptomycin (100 μg ml −1 ), L -glutamine (2 m M ) at 37°C in 5% CO 2 /95% air.

Techniques: Binding Assay

Journal:

Article Title: Pharmacological and biochemical characterization of adenosine receptors in the human malignant melanoma A375 cell line

doi: 10.1038/sj.bjp.0704352

Figure Lengend Snippet: Inhibition of [3H]-DPCPX (KH, KL, KI) binding by adenosine agonists and antagonists to human melanoma A375 membranes

Article Snippet: A375 cells (obtained from ATCC) were grown adherently and maintained in Dulbecco's Modified Eagle's Medium (DMEM), containing 10% foetal calf serum, penicillin (100 u ml −1 ), streptomycin (100 μg ml −1 ), L -glutamine (2 m M ) at 37°C in 5% CO 2 /95% air.

Techniques: Inhibition, Binding Assay

Journal:

Article Title: Pharmacological and biochemical characterization of adenosine receptors in the human malignant melanoma A375 cell line

doi: 10.1038/sj.bjp.0704352

Figure Lengend Snippet: Inhibition of [3H]-SCH 58261 binding (KI) by adenosine agonists and antagonists to human melanoma A375 membranes

Article Snippet: A375 cells (obtained from ATCC) were grown adherently and maintained in Dulbecco's Modified Eagle's Medium (DMEM), containing 10% foetal calf serum, penicillin (100 u ml −1 ), streptomycin (100 μg ml −1 ), L -glutamine (2 m M ) at 37°C in 5% CO 2 /95% air.

Techniques: Inhibition, Binding Assay

Journal:

Article Title: Pharmacological and biochemical characterization of adenosine receptors in the human malignant melanoma A375 cell line

doi: 10.1038/sj.bjp.0704352

Figure Lengend Snippet: Inhibition of [3H]-MRE 3008F20 (KH, KL, Ki) binding by adenosine agonists and antagonists to human melanoma A375 membranes

Article Snippet: A375 cells (obtained from ATCC) were grown adherently and maintained in Dulbecco's Modified Eagle's Medium (DMEM), containing 10% foetal calf serum, penicillin (100 u ml −1 ), streptomycin (100 μg ml −1 ), L -glutamine (2 m M ) at 37°C in 5% CO 2 /95% air.

Techniques: Inhibition, Binding Assay

Journal:

Article Title: Pharmacological and biochemical characterization of adenosine receptors in the human malignant melanoma A375 cell line

doi: 10.1038/sj.bjp.0704352

Figure Lengend Snippet: Stimulation curves of cyclic AMP levels by HE-NECA, NECA and CGS 21680 in A375 cells (A). Effect of increasing concentrations of NECA on cyclic AMP accumulation in A375 cells in the absence and in the presence of 100 nM A2A selective antagonist SCH 58261 (B). Data (mean of three experiments±s.e.mean) are given in percentages.

Article Snippet: A375 cells (obtained from ATCC) were grown adherently and maintained in Dulbecco's Modified Eagle's Medium (DMEM), containing 10% foetal calf serum, penicillin (100 u ml −1 ), streptomycin (100 μg ml −1 ), L -glutamine (2 m M ) at 37°C in 5% CO 2 /95% air.

Techniques:

Journal:

Article Title: Pharmacological and biochemical characterization of adenosine receptors in the human malignant melanoma A375 cell line

doi: 10.1038/sj.bjp.0704352

Figure Lengend Snippet: Effect of Cl-IB-MECA (30 μM) on intracellular calcium mobilization. Lower trace was obtained in calcium free medium and in the presence of 0.5 mM EGTA (A). Concentration dependent stimulation of Ca2+ mobilization by Cl-IB-MECA, IB-MECA and NECA in A375 cells (B). Data (mean of three experiments±s.e.mean) are given in percentages.

Article Snippet: A375 cells (obtained from ATCC) were grown adherently and maintained in Dulbecco's Modified Eagle's Medium (DMEM), containing 10% foetal calf serum, penicillin (100 u ml −1 ), streptomycin (100 μg ml −1 ), L -glutamine (2 m M ) at 37°C in 5% CO 2 /95% air.

Techniques: Concentration Assay

Journal:

Article Title: Pharmacological and biochemical characterization of adenosine receptors in the human malignant melanoma A375 cell line

doi: 10.1038/sj.bjp.0704352

Figure Lengend Snippet: Thermodynamic parameters observed for the binding of [3H]-DPCPX, [3H]-SCH 58261 and [3H]-MRE 3008F20 to A1, A2A and A3 receptors in A375 membranes, respectively

Article Snippet: A375 cells (obtained from ATCC) were grown adherently and maintained in Dulbecco's Modified Eagle's Medium (DMEM), containing 10% foetal calf serum, penicillin (100 u ml −1 ), streptomycin (100 μg ml −1 ), L -glutamine (2 m M ) at 37°C in 5% CO 2 /95% air.

Techniques: Binding Assay

Journal: The American journal of pathology

Article Title: The Neuropeptide Alpha-Melanocyte-Stimulating Hormone Is Critical for Corneal Endothelial Cell Protection and Graft Survival after Transplantation.

doi: 10.1016/j.ajpath.2021.10.016

Figure Lengend Snippet: Figure 1 Corneal endothelial cells (CEnC) express high levels of the receptor for a-MSH. A: Immunostaining for a-MSH receptor, melanocortin receptor 1 (MC1R), on murine (mCEnC) and human (hCEnC) CEnC lines and primary CEnCs. Blue: DAPI, purple: MC1R. B and C: Representative Western blot bands of MC1R in murine and human CEnC lines (B) and in primary murine corneal endothelium lysates (C). SIT lysate: small intestine tissue lysate used as positive control for mouse MC1R (mMC1R). A-375: human skin epithelial cell lysate used as positive control for human MC1R (hMC1R). Scale bars Z 50 mm.

Article Snippet: The mouse small intestine tissue (SIT) lysate (No. PK-AB718-1408; PromoCell, Heidelberg, Germany) and A375 human skin epithelial cell lysate (No. sc-3811; Santa Cruz Biotechnology, Dallas, TX) were used as positive controls for mouse and human MC1R, respectively.

Techniques: Immunostaining, Western Blot, Positive Control