Journal: Journal of proteome research

Article Title: Proteomic Analysis of Laser Microdissected Melanoma Cells from Skin Organ Cultures

doi: 10.1021/pr100164x

Figure Lengend Snippet: A. Cartoon depicting stages of melanoma progression in skin. Melanocytes reside in the epidermal/dermal junction. Early-stage radial growth phase melanoma cells descend into the papillary dermis and spread laterally. Invasion deep into the dermis characterizes melanomas in the vertical groth phase. B. Cartoon of a skin organ culture (SOC). SOCs are maintained at the air-liquid interface. The inset depicts a hematoxylin & eosin stained section collected on day 12 of culture.

Article Snippet: Normal human epidermal melanocytes (FC-0019, Lifeline Cell Technology, Walkersville, MD) were maintained in DermaLife Basal Medium with LifeFactors (LM-0027, Lifeline Cell Technology).

Techniques: Organ Culture, Staining

Journal: Journal of proteome research

Article Title: Proteomic Analysis of Laser Microdissected Melanoma Cells from Skin Organ Cultures

doi: 10.1021/pr100164x

Figure Lengend Snippet: A. Quantitative RT-PCR analysis of the expression of tenascin-C (TN-C) mRNA in WM983-A (VGP) and WM1158 (MGP) melanoma cells compared to human melanocytes, propagated in vitro. The data are expressed as the mean±standard deviation from three independent experiments. B. Immunoblot analysis depicting the level of TN-C in the VGP and MGP melanoma cell lines and melanocytes. C. Immunofluorescence analysis of melanocytes and VGP and MGP melanoma cell lines, probed with antibody to TN-C (pseudocolored green) and actin (pseudocolored red). The cells were counterstained with fluorescent DAPI (pseudocolored blue).

Article Snippet: Normal human epidermal melanocytes (FC-0019, Lifeline Cell Technology, Walkersville, MD) were maintained in DermaLife Basal Medium with LifeFactors (LM-0027, Lifeline Cell Technology).

Techniques: Quantitative RT-PCR, Expressing, In Vitro, Standard Deviation, Western Blot, Immunofluorescence

Journal: Journal of proteome research

Article Title: Proteomic Analysis of Laser Microdissected Melanoma Cells from Skin Organ Cultures

doi: 10.1021/pr100164x

Figure Lengend Snippet: A. Quantitative RT-PCR analysis of α-actinin-4 (ACN4) expression in the WM983-A (VGP) and WM1158 (MGP) melanoma cell lines and in melanocytes. The data are expressed as the mean ±standard deviation from three independent experiments. B. Immunoblot analysis of ACN4 expression in the VGP and MGP melanoma cell lines and in melanocytes. C. Immunofluorescence analysis of melanocytes and VGP and MGP melanoma cell lines, probed with antibody to ACN4 (pseudocolored green) and actin (pseudocolored red). The cells were counterstained with fluorescent DAPI (pseudocolored blue).

Article Snippet: Normal human epidermal melanocytes (FC-0019, Lifeline Cell Technology, Walkersville, MD) were maintained in DermaLife Basal Medium with LifeFactors (LM-0027, Lifeline Cell Technology).

Techniques: Quantitative RT-PCR, Expressing, Standard Deviation, Western Blot, Immunofluorescence

Journal: Oncotarget

Article Title: FOXP3 over-expression inhibits melanoma tumorigenesis via effects on proliferation and apoptosis.

doi:

Figure Lengend Snippet: (A) Distribution of FOXP3 expression in 146 stage III and IV melanomas. (B) Representative image of FOXP3 staining in a melanoma sample using the anti-FOXP3 Ab10563 antibody or IgG isotype control antibody (bottom right panel). Tumor cells with large irregular-shaped nuclei are indicated with red arrows while Treg cells with smaller, denser nuclei are indicated with green arrows. (C) Staining of melanomas and cultured melanocytes with anti-FOXP3 Ab20034 (clone 236A/E7) antibody. Sections were double-stained with anti-FOXP3 (brown) and anti-CD3 (Ferrangi blue) (left panel) or IgG isotype control antibodies (right panel) to distinguish between FOXP3 expressing tumor cells and infiltrating T cells. (D) Detection of FOXP3 mRNA expression in a panel of human melanoma cell lines using quantitative real-time PCR.

Article Snippet: Normal epidermal melanocyte cultures were obtained from Lonza Biosciences (Basel, Switzerland) and cultured according to the manufacturer's recommendation.

Techniques: Expressing, Staining, Control, Cell Culture, Real-time Polymerase Chain Reaction