As shown in Additional file 1: Figures S4 and S5, Iacs-miR-30C suppressed the β-catenin/Wnt signaling cascade in B16F10 tumor as support by the decreased protein level of Bcl9, β-catenin, C-myc and Cyclin D. Not surprisingly, compared with Anti-PD-1 or Iacs-miR-30C monotherapy, the combo treatment resulted in a significantly decreased amounts of regulatory T lymphocyte (CD4+/CD25+ cells, Fig. 6A, B) in sharp contrast to an increased number of tumor-infiltrating cytotoxic T lymphocyte (CD3+/CD8+ cells, Fig. 6C, D). This evidence concerns the gene PDCD1 and neoplasm.