FOXP3 and melanoma: However, considering that the proposed automatic gating identified a significant increase of only the FOXP3+ T cell subpopulation in melanoma patients, and that the manual gating did not produce any significant results on the analyzed subpopulations including non-Tregs and naive-Tregs, the immunological feature of melanoma patients most probably is in the cells with higher FOXP3 expressions, and the discordance in the FOXP3 boundary is probably not important in the setting.