Journal: Cancers

Article Title: Individualized Proteogenomics Reveals the Mutational Landscape of Melanoma Patients in Response to Immunotherapy

doi: 10.3390/cancers13215411

Figure Lengend Snippet: Comparison of tumor cells against melanocytes highlights patient-specific signaling pathways. ( A – D ) Scatter plot of log 2 -transformed ratios for proteins quantified in the PDX samples versus melanocytes for patient IDs 101 ( A ), 110 ( B ), 111 ( C ) and 129 ( D ). Significant proteins containing identified alternate peptides are marked in the respective color (significance B, p -value ≤ 0.05). Proteins marked in black were also identified in the corresponding FFPE material for each patient ID. The top 3 over-represented Reactome pathways based on all significantly up- or down regulated proteins are depicted in the upper part of each panel (Fisher-Exact test, p -value ≤ 0.2). ( E ) Heatmap of over-represented pathways within each patient based on proteins containing alternate variant peptides. Results are based on the Fisher-Exact test ( p -value ≤ 0.2). Color-coding indicates if a specific pathway was significantly over-represented in patient IDs 101 (blue), 110 (green), 111 (red) and 129 (orange).

Article Snippet: Primary human melanocytes and fibroblasts were isolated out of foreskin according to the protocol of CELLnTEC (CELLnTEC Advanced Cell Systems AG, Bern, Switzerland).

Techniques: Comparison, Protein-Protein interactions, Transformation Assay, Variant Assay

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Modulation of Diacylglycerol-Induced Melanogenesis in Human Melanoma and Primary Melanocytes: Role of Stress Chaperone Mortalin

doi: 10.1155/2019/9848969

Figure Lengend Snippet: Effect of OAG and TXC on melanin and tyrosinase contents in human melanoma (G361) (a) and primary melanocytes from Caucasian skin (PMC) (b). Cells were treated with OAG for 24 h followed by recovery either in control (OAG) or TXC-supplemented medium as indicated. Melanosome (c) and tyrosinase (d) immunofluorescence signals in control and OAG-treated cells, recovered in either control, or TXC-supplemented medium are shown. (e) Quantitation of the immunofluorescence for melanosome and tyrosinase are shown.

Article Snippet: Human skin melanoma (G361) cells and primary human melanocyte from Caucasian skin (PMC) were obtained from Japanese Collection of Research Bioresources (JCRB, Japan) and Kurabo Industries Ltd. (Osaka, Japan), respectively.

Techniques: Control, Immunofluorescence, Quantitation Assay

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Modulation of Diacylglycerol-Induced Melanogenesis in Human Melanoma and Primary Melanocytes: Role of Stress Chaperone Mortalin

doi: 10.1155/2019/9848969

Figure Lengend Snippet: Effect of OAG and TXC on mortalin, HSP70, and HSP60 expressions in human melanoma (G361) and primary melanocytes from Caucasian skin (PMC) as determined by immunocytochemistry using specific antibodies (a). Cells were treated with OAG for 24 h followed by recovery either in control (OAG) or TXC-supplemented medium as indicated. Effect of OAG and TXC on Reactive Oxygen Species (ROS) (b) and mitochondrial membrane potential (c) in human melanoma (G361) and primary melanocytes from Caucasian skin (PMC) as determined by live cell ROS detection and JCI staining, respectively. Cells were treated with OAG for 24 h followed by recovery either in the control (OAG) or TXC-supplemented medium as indicated.

Article Snippet: Human skin melanoma (G361) cells and primary human melanocyte from Caucasian skin (PMC) were obtained from Japanese Collection of Research Bioresources (JCRB, Japan) and Kurabo Industries Ltd. (Osaka, Japan), respectively.

Techniques: Immunocytochemistry, Control, Membrane, Staining

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Modulation of Diacylglycerol-Induced Melanogenesis in Human Melanoma and Primary Melanocytes: Role of Stress Chaperone Mortalin

doi: 10.1155/2019/9848969

Figure Lengend Snippet: Effect of selected natural extracts on OAG-induced increase in melanosomes (a) and melanin content (b) in human melanoma (G361) and primary melanocytes from Caucasian skin (PMC). Cells were treated with OAG for 24 h followed by recovery either in control (OAG) or extract (EX-33, EX-39, EX-45, and EX-46) supplemented medium as indicated. Increase in OAG-induced ROS and its reversal in the presence of extracts are shown (c). Increase in melanosomes, Tyrosinase, HSP60, and mortalin in OAG-treated human melanoma (G361) (d) and primary melanocytes from Caucasian skin (PMC) (e) and their reversal in response to treatment with extract (EX-33, EX-39, EX-45, and EX-46) are shown. Quantitation of the data obtained from three independent experiments is shown with each panel.

Article Snippet: Human skin melanoma (G361) cells and primary human melanocyte from Caucasian skin (PMC) were obtained from Japanese Collection of Research Bioresources (JCRB, Japan) and Kurabo Industries Ltd. (Osaka, Japan), respectively.

Techniques: Control, Quantitation Assay

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Modulation of Diacylglycerol-Induced Melanogenesis in Human Melanoma and Primary Melanocytes: Role of Stress Chaperone Mortalin

doi: 10.1155/2019/9848969

Figure Lengend Snippet: Effect of selected natural extracts on OAG-induced decrease in mitochondrial membrane potential and their reversal in the presence of TXC and extracts in both human melanoma (G361) and primary melanocytes (PMC) is shown.

Article Snippet: Human skin melanoma (G361) cells and primary human melanocyte from Caucasian skin (PMC) were obtained from Japanese Collection of Research Bioresources (JCRB, Japan) and Kurabo Industries Ltd. (Osaka, Japan), respectively.

Techniques: Membrane

Journal: Experimental dermatology

Article Title: Divergence of cAMP signaling pathways mediating augmented nucleotide excision repair and pigment induction in melanocytes

doi: 10.1111/exd.13291

Figure Lengend Snippet: (A,B) SK-MEL-2 melanoma cells (n=3) (A,C) and primary human melanocytes (PHMs; n=4) (B,D) were treated with scrambled siRNA or siRNA directed to MITF prior to treatment with 10 uM forskolin. Whole cell lysates were collected at 1 hour and pSer435-ATR levels were determined by kinase assay (A,B). Values not sharing a common letter are significantly different as determined by one-way ANOVA and Tukey post-hoc test (p<0.05). Data are expressed as mean-fold change over control ± SEM. C,D) Cells were treated with scrambled siRNA or siRNA directed to MITF. Cells were treated with 10 uM forskolin for 30 minutes prior to treatment with 10 J/m2 UVB radiation. MITF knockdown following treatment with siRNA directed to MITF is shown in inset. Significance between control and forskolin treatment as determined by two-way paired ANOVA (p<0.05) is denoted by an asterisk (*). Data are expressed as mean CPD remaining ± SEM. Insets show degree of MITF knockdown by Western blotting.

Article Snippet: Transformed melanoma SK-MEL-2 (ATCC) cells and primary human melanocytes (Coriell Institute for Medical Research) were cultured in Roswell Park Memorial Institute (RMPI) media (Life Technologies) with 10% fetal bovine serum and Cascade Biologics Medium 254 (Life Technologies) respectively.

Techniques: Kinase Assay, Control, Knockdown, Western Blot

Journal: Experimental dermatology

Article Title: Divergence of cAMP signaling pathways mediating augmented nucleotide excision repair and pigment induction in melanocytes

doi: 10.1111/exd.13291

Figure Lengend Snippet: SK-MEL-2 melanoma cells (n=3) (A,C) and primary human melanocytes (PHMs; n=4) (B,D) were treated with vehicle controls 10 uM VE-821 or 10 uM forskolin. Cells were collected, incubated with L-DOPA (see methods) and pelleted after 2h. A,B) photographs of microcentrifuge tubes showing cell pellets of indicated conditions. C,D) Cells were lysed in soluene-350 solution and spectrophotometry of supernatants was quantified at 492 nm. Values not sharing a common letter are significantly different as determined by one-way ANOVA and Tukey post hoc test (p<0.05). Data are expressed as mean-fold change over control ± SEM.

Article Snippet: Transformed melanoma SK-MEL-2 (ATCC) cells and primary human melanocytes (Coriell Institute for Medical Research) were cultured in Roswell Park Memorial Institute (RMPI) media (Life Technologies) with 10% fetal bovine serum and Cascade Biologics Medium 254 (Life Technologies) respectively.

Techniques: Incubation, Spectrophotometry, Control

Journal:

Article Title: Gpnmb is a melanosome-associated glycoprotein that contributes to melanocyte/keratinocyte adhesion in a RGD-dependent fashion

doi: 10.1111/j.1600-0625.2008.00830.x

Figure Lengend Snippet: (A) Human Gpnmb-Fc, mouse IgG (2 µg/lane), or whole cell extracts (40 µg/lane) from COS-1 transfected with vector or phGpnmb vector (encoding human Gpnmb), or 293T cells were examined by immunoblotting for immunoreactivity to 3D5 mouse anti-human Gpnmb mAb. (B) Protein expression of Gpnmb in SK-MEL-28 cells was compared with that in primary-cultured human normal melanocytes. Expression of β-actin was also examined. (C) Mouse Gpnmb-Fc, human IgG, or whole cell extracts (40 µg/lane) from COS-1 cells transfected with pmGpnmb (encoding mouse Gpnmb) or empty vector, EL-4 or B16F10 cells were also subjected to immunoblotting using UTX-103 rabbit anti-mouse Gpnmb mAb. Protein bands immunoreactive to 3D5 or UTX-103 mAb are shown by closed arrowheads. Open arrowhead indicates non-specific band. (D) Whole cell extract from SK-MEL-28 or B16F10 was treated with (+)/without (−) N-glycosidase (N-Gly), followed by immunoblotting using anti-human (3D5) or anti-mouse Gpnmb mAb (UTX-103 or 1E4). (E) P815 cells transfected with pmGpnmb or vector alone were examined by flow-cytometry for surface expression of Gpnmb using UTX-103 mAb. B16F10 melanoma cells were left untreated (for surface expression) or permealized (for intracellular expression) and stained with UTX-103 mAb (open histogram) or control IgG (filled in gray). Expression was examined by flow-cytometry.

Article Snippet: Human primary melanocytes were purchased from Cambrex Bio Science Walkersville Inc. (Walkersville, MD), and maintained in Melanocyte Cell Basal Medium-4 and MGM-4 Single Quots (both from Cambrex).

Techniques: Transfection, Plasmid Preparation, Western Blot, Expressing, Cell Culture, Flow Cytometry, Staining, Control

Journal:

Article Title: Gpnmb is a melanosome-associated glycoprotein that contributes to melanocyte/keratinocyte adhesion in a RGD-dependent fashion

doi: 10.1111/j.1600-0625.2008.00830.x

Figure Lengend Snippet: Human melanocytes were cultured in the presence of different agents. After culturing for 1 day, cells were stained with 3D5 mAb (open histograms) or control IgG (closed histograms). Staining was examined by flow-cytometry. Mean fluorescence intensity (MFI) of staining by 3D5 mAb is shown in the histograms. Second experiment showed similar results.

Article Snippet: Human primary melanocytes were purchased from Cambrex Bio Science Walkersville Inc. (Walkersville, MD), and maintained in Melanocyte Cell Basal Medium-4 and MGM-4 Single Quots (both from Cambrex).

Techniques: Cell Culture, Staining, Control, Flow Cytometry, Fluorescence