Seven days following tumor cell injection, mice were randomly divided into 6 groups (9–10 mice per group) to receive 1 of the following 6 therapies: (i) CH/Endo28-control miR inhibitor, (ii) CH/Endo28-control miR inhibitor + paclitaxel, (iii) CH/Endo28-miR106b-5p inhibitor, (iv) CH/Endo28-miR106b-5p inhibitor + paclitaxel, (v) CH/Endo28-miR30c-5p inhibitor, or (vi) CH/Endo28-miR30c-5p inhibitor + paclitaxel. Mice were sacrificed when any animals in a control or treatment group became moribund (after 3 to 4 weeks of therapy). (A) Mean tumor weight. (B) Mean number of tumor nodules. (C) Representative images of tumor burden from at least 5 mice per group. (D) Representative tumor sections showing effect of miR silencing on pericyte coverage. Tumor sections were stained with anti-desmin (pericyte coverage marker) and anti-CD31 (for blood vessels) antibodies respectively (red indicates blood vessels, green indicates pericyte coverage, and blue indicates nuclei). (E) Representative tumor sections showing effect of miR silencing on vascular permeability of blood vessels after i.v. injection of FITC-dextran. Tumor sections were stained with anti-CD31 antibody (red indicates blood vessels, green indicates FITC-dextran, and blue indicates nuclei). In D and E, staining was done on 5 sections per group. (F) Determination of extent of hypoxia after staining tumor sections with hypoxia marker anti-CA9 and anti-CD31 antibodies (red indicates blood vessels, green indicates hypoxia, and blue indicates nuclei). (G) Quantification of pericyte coverage and extravasated FITC-dextran. Pericyte coverage was determined by the percentage of vessels with ≥ 50% coverage of desmin in positive cells in 5 random fields of each section at ×200 magnification of each tumor. Quantification of extravasated FITC-dextran was carried out using fluorescence microscopy under the green fluorescent filter using the following scoring system: 0 points, no staining; 1 point, <25%; 2 points, 25%–50%; 3 points, 50%–75%; 4 points, 75%–100% FITC-dextran (using 8–10 sections per tumor at ×200 magnification. (H) Quantification of average number of blood vessels after indicated treatments. To quantify microvessel density, we recorded the number of blood vessels that stained positive for CD31 in 5 random fields of each section at ×200 magnification for each sample. Values are the mean ± SEM. *P < 0.05, **P < 0.01, (1-way ANOVA followed by a Tukey’s multiple comparison post-hoc test). NS, not significant. Scale bars: 100 μm. CH., chitosan; miR-inh, miRNA inhibitor.