Journal: Theranostics

Article Title: Heterogeneity of tyrosine-based melanin anabolism regulates pulmonary and cerebral organotropic colonization microenvironment of melanoma cells

doi: 10.7150/thno.69198

Figure Lengend Snippet: Tyrosine-induced melanosome -dependent cell death supported the acquisition of tumor-promoting phenotype by macrophages. A. Relative cell viability was evaluated by MTS assay. The same number of B16 cells were cultured in the conditional medium with different tyrosine addition for 24 h. B-C. Cell apoptosis was examined through Annexin V / PI staining. B16 cells were treated with different concentration of tyrosine or ch282-5 (Bcl-2 inhibitor, positive control). D-E. Cell cycle was analyzed by PI staining. F-G. Mitochondria was quantized by Mito-tracker staining. H-J. Mitochondrial membrane potential (MMP) dependent TMRE signaling was detected by flow cytometry (H-I). MMP was reflected by the MFI ratio of TMRE to Mito-tracker (J). K-L. Lysosome were quantized by Lyso-tracker staining. M. Representative photos of Tyr-L cells and Tyr-H cells with 10 T tyrosine for 24 h. N-O. Representative photos of B16 cells. B16 cells with 10 T tyrosine combining with or without deoxyarbutin or hydroquinone for 48 h. The percentage of pigment cells were calculated. P. Peritoneal macrophages were co-cultured with the EMCDRs (labeled with ZsGreen) in different ratios. Consuming efficiency were identified by flow cytometry. Q-R. Photos of the lung with metastasis lesions from experimental pulmonary metastasis model (Q). Normal B16 cells were injected intravenously, and mice were treated with PBS, macrophages, or EMCDRs-educated macrophages respectively. Percent survival was calculated (R). Mφ, macrophages; EMCDRs-Mφ, EMCDRs educated macrophages. All data given as mean ± SEM., n ≥ 3. *P < 0.05 , **P < 0.01 , ***P < 0.001 , ns, not significant. All results shown are representative of three independent experiments.

Article Snippet: Hydroquinone (Selleck, S4580) at the dosage of 50 mg/kg.

Techniques: MTS Assay, Cell Culture, Staining, Concentration Assay, Positive Control, Membrane, Flow Cytometry, Labeling, Injection

Journal: Theranostics

Article Title: Heterogeneity of tyrosine-based melanin anabolism regulates pulmonary and cerebral organotropic colonization microenvironment of melanoma cells

doi: 10.7150/thno.69198

Figure Lengend Snippet: Targeting tyrosine utilization augments the therapeutic effects of PD1 monoclonal antibody. A. The schedule showing composite melanoma model and treatment. B-F. Composite melanoma mice were treated with E.T.G. (2-Ethoxybenzamide, tyrosine, plus GSK343), PD1 monoclonal antibody (mAb), and E.T.G. plus PD1 mAb, PBS treatment as a control. Percent survival (B), in situ tumor mass (C), pulmonary macrometastases (D), cerebral colonization (E) and cerebral tumor weight (F) were observed. G-K. Composite melanoma mice were treated with hydroquinone plus CXCR2 inhibitor SB225002 (Hy + CXCR2i), PD1 mAb, and Hy + CXCR2i plus PD1 mAb, PBS treatment as a control. The indicators as panel B-F were observed. All data given as mean ± SEM., n ≥ 3. *P < 0.05 , ***P < 0.001 , ns, not significant. All results shown are representative of three independent experiments.

Article Snippet: Hydroquinone (Selleck, S4580) at the dosage of 50 mg/kg.

Techniques: Control, In Situ