Journal: Melanoma Research

Article Title: Interfering with aggregated α-synuclein in advanced melanoma leads to a major upregulation of MHC class II proteins

doi: 10.1097/CMR.0000000000000982

Figure Lengend Snippet: Upregulation of MHC class II antigen in human melanoma xenografts treated systemically with anle138b, and expression of α-synuclein, MHC class II, and MHC class I proteins in melanoma TMA cores. (a) A tissue section from a nontreated WM983-B tumor xenograft probed with an antibody to human MHC class II (HLA-DPB1) protein (pseudocolored green). (b) A tissue section from an anle138b-treated WM983-B tumor xenograft probed with an antibody to human MHC class II (HLA-DPB1) protein (pseudocolored green). (c) Fluorescence intensity level of expression of proteins – α-synuclein (pseudocolored red), MHC II (pseudocolored yellow), MHC I (pseudocolored green) – in areas, encircled by a white line, in tissue sections from three of the Stage II and from one of the Stage IV melanoma TMA cores in which α-synuclein or vice versa, MHC II, or MHC I protein was expressed more strongly. TMA, tissue microarray.

Article Snippet: Four melanoma tissue microarray (TMA) slides ( https://www.biomax.us/tissue-arrays/Melanoma/ME811 ), each containing two tissue sections from the same 39 cases of melanoma, and tissue sections from three cases of normal skin tissue were heated at 60 °C, deparaffinized and rehydrated.

Techniques: Expressing, Fluorescence, Microarray

Journal: PLoS ONE

Article Title: Coordinate Autophagy and mTOR Pathway Inhibition Enhances Cell Death in Melanoma

doi: 10.1371/journal.pone.0055096

Figure Lengend Snippet: (A) Melanoma cell lines were transiently transfected with an EGFP-LC3 expressing plasmid and grown in normal medium. 24 hours after transfection, the cells were fixed and the autophagosomes in the cells were visualized by the presence of LC3 puncta under fluorescence microscopy, with the percentages of cells showing LC3 puncta (mean ± SD) indicated. All tested cell lines showed positive punctation indicating high basal autophagy. (B) Western blot shows that melanoma cell lines have high basal LC3-II and p62 protein levels. Upon HBSS induced starvation, increased cleavage of LC3-I to LC3-II indicative of autophagy induction and decreased or unchanged p62 levels were seen. Extracts from autophagy competent and autophagy deficient iBMK cells served as positive and negative controls. (C) Immunofluorescence staining for endogenous LC3 on a human melanoma tissue microarray. Punctate LC3 localization for autophagosomes is indicated by red arrows (top panel). The percentages of specimens with punctate LC3 staining in malignant, metastatic and benign nevus groups are indicated in the table (bottom panel). (D) Western blot showed decreased expression levels of Atg7 (top panel) and impaired clonogenic survival (bottom panel) in response to lentiviral shRNA knockdown of the essential autophagy regulator Atg7.

Article Snippet: Human Malignant Melanoma Tissue Microarray (US Biomax, Inc) slides, or xenograft tumor paraffin slides were deparaffinized, hydrated and incubated with primary antibody against LC3 (Nanotools, 1∶100 dilution) for 15 min at room temperature followed by FITC tagged anti-mouse (Jackson Immuno Research) antibody, then mounted and visualized by epifluorescence microscopy.

Techniques: Transfection, Expressing, Plasmid Preparation, Fluorescence, Microscopy, Western Blot, Immunofluorescence, Staining, Microarray, shRNA

Journal: Scientific Reports

Article Title: Yes-Associated Protein (YAP) Promotes Tumorigenesis in Melanoma Cells Through Stimulation of Low-Density Lipoprotein Receptor-Related Protein 1 (LRP1)

doi: 10.1038/s41598-017-14764-4

Figure Lengend Snippet: The consistency of YAP and LRP1 in tissue microarray specimen. ( a , b ) TMA slides include forty skin melanoma tissues and eight skin normal tissues which locate on the bottom of the each TMA. Representative images of IHC from HCC TMA stained with anti-YAP or anti-LRP1 antibodies. Scale bar, 100 μM. ( c ) Representative images of IHC from skin melanoma HCC TMA stained with anti-YAP or anti-LRP1 antibodies. Scale bar, 100 μM. ( d ) The statistical figure of skin melanoma IHC images from HCC TMA stained with anti-YAP or anti-LRP1 antibodies. The TMA data were analyzed using the χ2 test.

Article Snippet: For IHC, melanoma tissue microarray analysis (TMA) slides were purchased from Biomax (Rockville, MD, USA).

Techniques: Microarray, Staining