Fig. 1

Effectiveness of IL-10 silencing in vitro and in vivo. a Scheme of lentiviral vector encoding shRNA against murine IL-10 or scrambled sequence against GAPDH (shN) and selected shRNA sequences used in experiments; b Relative expression of IL-10 mRNA in transduced BMDC measured using Real-Time PCR; c EGFP expression in BMDC on the 2nd day after transduction with LVs as a control of the transduction effectiveness; d Concentration of IL-10 in supernatants collected after 24 h culture of shIL10 transduced and control BMDC measured using ELISA. The results are given as a mean ± SD calculated for three repeats in three independent experiments; e-j Intratumoral activity of shIL10–3 LVs. MC38 tumors were dissected on the 6th day after triple intratumoral injection of LVs encoding shIL10–3 or shN; e IL-10 produced by cells isolated from tumors; f Differences in EGFP expression between tumor cells, and lymphocytes or myeloid cells infiltrating tumor; g EGFP expression in defined subpopulations of myeloid cells; h, i The percentage of selected lymphocytic or myeloid cell subpopulations among leukocytes infiltrating into tumor after LVs injection; j MHC II expression in defined subpopulations of myeloid cells. To calculate the mean ± SD, at least six mice per group were analyzed. The differences between the groups were estimated using non-parametric Kruskal-Wallis test followed by Dunn’s multi comparison test (* p < 0.05, ** p < 0.01, *** p < 0.001)