Journal: Pigment cell & melanoma research

Article Title: p53 prevents progression of nevi to melanoma predominantly through cell cycle regulation

doi: 10.1111/j.1755-148X.2010.00773.x

Figure Lengend Snippet: High p53 delays melanoma formation but primarily suppresses the progression of primary to metastatic melanoma. (A) Scatter plot of age of onset of 1st pigmented lesion (PL) (n = 25 for TP-ras0/+ and n = 11 for TP-ras0/+: Mdm4+/− mice), melanomas (n = 14 for TP-ras0/+ and n = 11 for TP-ras0/+: Mdm4+/− mice), and the age of onset of all individual pigmented lesions (n = 113 for TP-ras0/+ and n = 47 for TP-ras0/+: Mdm4+/− mice). The mean is represented by the bar in the center of each plotted data series. ns, not significant (P > 0.05); 1 star, significant (P = 0.01–0.05); 2 stars, very significant (P = 0.001–0.01). (B) Histogram presenting the percentage of mice with pigmented lesions (PL) classified by tumor size and progression. The labels for each of the 5, 10, and 20 mm2 groupings refer to tumors that are ≥ the indicated size.

Article Snippet: The antibodies used were rabbit p53 (CM5, 1:400 dilution; Vector Laboratories Inc., Burlingame, CA, USA), mouse S100 (1:5000 dilution; Abcam, Cambridge, MA, USA) and rabbit Ki67 (1:1000; Vector Laboratories Inc.).

Techniques:

Journal: Pigment cell & melanoma research

Article Title: p53 prevents progression of nevi to melanoma predominantly through cell cycle regulation

doi: 10.1111/j.1755-148X.2010.00773.x

Figure Lengend Snippet: High p53 prohibited tumor growth. Growth (mm2) of individual tumors of TP-ras0/+ mice (A) and TP-ras0/+: Mdm4+/− mice (B) followed every 10 days from emergence. (C) Box and whisker plot of growth rate (mm2/day) of these tumors based on histological presentation (nevus or melanoma) and size (<5, 5–10 and >10 mm2). The whiskers represent the minimum and maximum growth rates. (D) Kaplan–Meier presentation of the survival of TP-ras0/+ (n = 28) and TP-ras0/+: Mdm4+/− mice (n = 17), Log-rank test P = 0.0441.

Article Snippet: The antibodies used were rabbit p53 (CM5, 1:400 dilution; Vector Laboratories Inc., Burlingame, CA, USA), mouse S100 (1:5000 dilution; Abcam, Cambridge, MA, USA) and rabbit Ki67 (1:1000; Vector Laboratories Inc.).

Techniques: Whisker Assay

Journal: Pigment cell & melanoma research

Article Title: p53 prevents progression of nevi to melanoma predominantly through cell cycle regulation

doi: 10.1111/j.1755-148X.2010.00773.x

Figure Lengend Snippet: Histological characterization of pigmented lesions. (A) Scatter plot of p53 immunopositive cells in 25 TP-ras0/+ nevi, 12 TP-ras0/+ melanomas, 13 TP-ras0/+: Mdm4+/− nevi, and 4 TP-ras0/+: Mdm4+/− melanomas (>5 mm2) analyzed per field (0.5 mm2 area using 20× magnification). (B) Plot of tumor progression (based on size) in relation to degree of pigmentation of each pigmented lesion using the reference scale from 1 (highest, 100%) to 5 (lowest, 0–10%). A representative picture of hematoxylin and eosin (HE) sections of pigmented lesion scored by the degree of pigmentation (at the bottom).

Article Snippet: The antibodies used were rabbit p53 (CM5, 1:400 dilution; Vector Laboratories Inc., Burlingame, CA, USA), mouse S100 (1:5000 dilution; Abcam, Cambridge, MA, USA) and rabbit Ki67 (1:1000; Vector Laboratories Inc.).

Techniques:

Journal: Pigment cell & melanoma research

Article Title: p53 prevents progression of nevi to melanoma predominantly through cell cycle regulation

doi: 10.1111/j.1755-148X.2010.00773.x

Figure Lengend Snippet: Effect of Nutlin-3 on clonogenecity of melanoma cell lines. (A) A representative picture (4× magnification) of colony-forming assay of four cell lines (BL: mutant p53, A04 and MM329: wild-type p53). Cell lines treated (NT) or treated with (10, 30 μM) Nutlin-3 were plated on Matrigel matrix and allowed to grow for 72 h before colony counting. All colonies ≥50 cells in the chamber were counted. (B) A graphic representation of the effect of Nutlin-3 treatment on colony counts of tested cell lines.

Article Snippet: The antibodies used were rabbit p53 (CM5, 1:400 dilution; Vector Laboratories Inc., Burlingame, CA, USA), mouse S100 (1:5000 dilution; Abcam, Cambridge, MA, USA) and rabbit Ki67 (1:1000; Vector Laboratories Inc.).

Techniques: Mutagenesis

Journal: Pigment cell & melanoma research

Article Title: p53 prevents progression of nevi to melanoma predominantly through cell cycle regulation

doi: 10.1111/j.1755-148X.2010.00773.x

Figure Lengend Snippet: Analysis of p53 target genes in melanocytes treated with Nutlin-3 for 18 h. (A) Representation of p53 target genes based on microarray results. In red, up-regulated genes; in white, genes with no differential expression; in blue, down-regulated genes. (B) Representation of up-regulated (red) and down-regulated (blue) genes in the CDKN1A network generated by Ingenuity pathway analysis tools. Arrows represent the regulatory relationships between genes. (C) Schematic representation of tumor-suppressive activity of p53 in Ras-dependent DMBA-induced progression model from a melanocyte to nevus to metastatic melanoma. GA, growth arrest.

Article Snippet: The antibodies used were rabbit p53 (CM5, 1:400 dilution; Vector Laboratories Inc., Burlingame, CA, USA), mouse S100 (1:5000 dilution; Abcam, Cambridge, MA, USA) and rabbit Ki67 (1:1000; Vector Laboratories Inc.).

Techniques: Microarray, Expressing, Generated, Activity Assay

Journal: Biomicrofluidics

Article Title: On-chip immuno-agglutination assay based on a dynamic magnetic bead clump and a sheath-less flow cytometry

doi: 10.1063/1.5093766

Figure Lengend Snippet: Dose-response curve of the antigen/antibody assay for IgG. (a) Percentage of monomers vs sample concentration of both rabbit IgG and negative control and (b) normalized signal vs concentration of rabbit IgG. Each dot is the average result of five measurements, and the error bars represent SD.

Article Snippet: In the second set of experiments, goat anti-rabbit IgG coated MBs are prepared by adding protein A-coated MBs suspension of 5 μ l (Dynabeads Protein A for Immunoprecipitation, ThermoFisher Scientific Co., USA) to 10 μ g goat anti-rabbit IgG (bs-0295G, Bioss Antibodies Co., China) diluted in 40 μ l PBS with 0.1% Tween-20, followed by surface-sealing with goat serum (C-0005, Bioss Antibodies Co., China) of 50 μ l. The processed rabbit IgG (bs-0295P, Bioss Antibodies Co., China) is then diluted by factors of 103–108 with PBS and used as analyte.

Techniques: Concentration Assay, Negative Control