Fig. 4

miR-130a & miR-145 reprogram Gr-1⁺CD11b⁺ cells and enhance host immunity. a TβRII Western blot (left) and qRT-PCR (right) of Gr-1⁺CD11b⁺ cells from 4T1 tumor-bearing mice 24 h after miR control, miR-130a and miR-145 electroporation. b qRT-PCR of TβRII after electroporation of miR-130a or miR-145 with or without miRNA antisense inhibitors. c M1/M2 cytokine ratio post treatment with miRNA electroporation with or without miR-130a or miR-145 antisense inhibitors (n = 5). The ratio of M1/M2 cytokines was calculated by dividing each M1 cytokine (TNFα, IL-12, GM-CSF) to M2 cytokine (IL-10, TGFβ1, IL-4) as described in Material and Methods. d Left panels: flow cytometry of IFNγ⁺CD8⁺ T cells from splenocytes of TCR-HA transgenic mice co-cultured with Gr-1⁺CD11b⁺ myeloid cells with or without miRNA, which were pulsed with HA peptide. Right panels: quantitative data. The Gr-1⁺CD11b⁺ cells (Mye) to splenocytes (Spl) ratio is indicated for each panel. e IFNγELISPOT of splenocytes from TCR-HA transgenic mice co-cultured with Gr-1⁺CD11b⁺ cells following miRNA electroporation. f CTL assays: flow cytometry of B16 melanoma tumor cells (Sytox blue positive are dead) expressing hgp100 co-cultured with splenocytes from Pmel-1 transgenic mice and Gr-1⁺CD11b⁺ cells from spleens of miR-130a transgenic, quantitative data on right. The percentage of targeted dead tumor cells = total dead cells + subtraction of dead cells from a single culture of tumor cells, myeloid cells, and splenocytes. All data are presented as mean±SEM, and Student’s t test was performed. *p < 0.05, **p < 0.01, ***p < 0.001