Journal: Cell

Article Title: Single-cell Map of Diverse Immune Phenotypes in the Breast Tumor Microenvironment

doi: 10.1016/j.cell.2018.05.060

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: anti-TIM3 154Sm , Fluidigm , Cat:3154010B.

Techniques: Staining, Antibody Labeling, Software, Diffusion-based Assay

Journal: The Journal of investigative dermatology

Article Title: TIM-3 is expressed in melanoma cells and is upregulated in TGF-beta stimulated mast cells.

doi: 10.1038/sj.jid.5700616

Figure Lengend Snippet: Figure 4. TIM-3 is expressed both in melanoma-surrounding human mast cells and in melanoma cells. (a) Immunohistochemistry with anti-TIM3 antibody in melanoma tissue sections. Two representatives out of three different patients. The background staining of the control slides was negligible (see Materials and Methods). Single asterisks indicate melanoma cells. Double asterisk labels the area that is magnified in (b). Bar ¼ 20 mm. (b) Asterisks mark mast cells in serial sections. Left side: immunohistochemistry with anti-TIM3 antibody (the magnification from a part of a). Right side: mast cells show metachromatic staining with toluidine blue. Bar ¼ 20 mm. (c) WM35 and HT168-M1 melanoma cell lines express TIM-3 as detected by real-time PCR with TaqMan probes fitting to either exon6/7 or exon2/3 (mean þ SEM, n ¼ 4, the WM35 data obtained by TaqMan assay fitting to exon 2/3 were taken as 100%). (d) Both WM35 and HT168-M1 melanoma cell lines were positive for cell surface TIM-3 (flow cytometric analysis). Simple line: isotype control, bold line: anti-TIM3. (e) Melanoma cell lines express TIM-3 at a higher level than HEMn-DP melanocytes (flow cytometry, simple line: melanocytes; bold line: WM35; dotted line: isotype control. With HT168-M1 similar results were obtained).

Article Snippet: The following antibodies were used: goat anti-human cathepsin G (0.5 mg/ml), goat anti-human Tim3 (0.5 mg/ml), goat anti-human GDF15 (0.5 mg/ml) (R&D Systems), rat anti-tubulin (0.5 mg/ml, Serotec), anti-rat horseradish peroxidase (1:10,000), anti-goat horse- radish peroxidase (1:8,000, Sigma).

Techniques: Immunohistochemistry, Staining, Control, Real-time Polymerase Chain Reaction, TaqMan Assay, Flow Cytometry