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. 2023 Aug 9;13(10):2270–2291. doi: 10.1158/2159-8290.CD-23-0389

Figure 6.

Figure 6. GABA-A receptor activation drives melanoma growth in mouse models and human 3D skin reconstructs. A, Schematic representation of the mouse ear transplant experiment in which YUMM 1.1 cells expressing control-GFP or Gad1-GFP are injected into mouse ears and imaged at 4, 8, and 12 days after injection. Created using BioRender.com. B, Representative images of mouse ears transplanted with control- or Gad1-GFP–expressing YUMM 1.1 mouse melanoma cells, 4 days after injection. C, Mean fluorescence intensity (MFI) in control- or Gad1-GFP–expressing mouse melanoma cells transplanted into mouse ears and imaged at 4, 8, and 12 days after injection in 4 biological replicates. Error bars, SD; P values generated by two-tailed unpaired t test; *, P < 0.05; **, P < 0.01. D, Schematic representation of 3D melanoma skin reconstruction experiments, in which the constructs were treated with the GABA-A receptor agonist muscimol. Created using BioRender.com. E, Representative images of IHC performed on 3D melanoma skin reconstructs treated with control media or muscimol-containing media, with melanoma cells stained with an anti-BRAFV600E antibody, after 24 days in coculture with keratinocytes and fibroblasts in 3D reconstruct media. F, Percentage melanoma-positive area (BRAFV600E-positive) in 3D skin reconstructs treated with control media or muscimol-containing media (100 μmol/L). Data are pooled from 3 biological replicates, normalized to control media. Error bars, SD; P values generated by two-tailed unpaired t test; **, P < 0.005.

GABA-A receptor activation drives melanoma growth in mouse models and human 3D skin reconstructs. A, Schematic representation of the mouse ear transplant experiment in which YUMM 1.1 cells expressing control-GFP or Gad1-GFP are injected into mouse ears and imaged at 4, 8, and 12 days after injection. Created using BioRender.com. B, Representative images of mouse ears transplanted with control- or Gad1-GFP–expressing YUMM 1.1 mouse melanoma cells, 4 days after injection. C, Mean fluorescence intensity (MFI) in control- or Gad1-GFP–expressing mouse melanoma cells transplanted into mouse ears and imaged at 4, 8, and 12 days after injection in 4 biological replicates. Error bars, SD; P values generated by two-tailed unpaired t test; *, P < 0.05; **, P < 0.01. D, Schematic representation of 3D melanoma skin reconstruction experiments, in which the constructs were treated with the GABA-A receptor agonist muscimol. Created using BioRender.com. E, Representative images of IHC performed on 3D melanoma skin reconstructs treated with control media or muscimol-containing media, with melanoma cells stained with an anti-BRAFV600E antibody, after 24 days in coculture with keratinocytes and fibroblasts in 3D reconstruct media. F, Percentage melanoma-positive area (BRAFV600E-positive) in 3D skin reconstructs treated with control media or muscimol-containing media (100 μmol/L). Data are pooled from 3 biological replicates, normalized to control media. Error bars, SD; P values generated by two-tailed unpaired t test; **, P < 0.005.