Figure 4.
Notes: (A) Untreated control (three mice), (B) PLGA-empty (four mice), and (C) PLGA-OP (four mice) 20 mg treated cohorts. Necropsy tumors, HE staining of tumors, and paraffin-embedded tumor sections (5 μm) on glass slides were processed for immunohistochemistry using primary DyLight 488 conjugated rat monoclonal anti-mouse CD31+ (PECAM-1) antibody, primary anti-E-cadherin, and N-cadherin antibodies followed with polyclonal goat anti-rabbit Alexa Fluor® 488 secondary antibody and Entellan® rapid mounting media. Background control (not shown) sections were prepared without the primary antibodies and relative staining density was 2–4×105. The bar on the stained tissue sections represents 200 μm. Images are representative of at least five fields of view from two tumor sections. (D) Quantitative analysis was done by assessing the density of tumor staining corrected for background in each panel using Corel Photo Paint 8.0 software. Each symbol in the figure represents the mean ± SEM corrected density of tumor staining within the respective images. Statistical analysis using unpaired t-test was carried out using GraphPad Prism and results were compared with the untreated cohort.
Abbreviations: OP, oseltamivir phosphate; PLGA, poly (lactic-co-glycolic acid); SEM, standard error of the mean.