Fig. 3.
Human U87MG and T98G glioma cell treatment in vitro with nanoconjugates containing pH-dependent or pH-independent endosome escape units. (A) Effects of LLL and LOEt endosomal escape units on U87MG cell viability. Arrows relate to microscopic views (20× magnification) after 24 h treatment with 0.5 mg of either nanoconjugate. The cells treated with P/LOEt unlike P/LLL had low viability at high concentrations (Left) and were in early apoptosis (Center). (Right) Representative FACS analysis of cell death after double staining of cells with propidium iodide and FITC Annexin V. Note markedly increased fraction of apoptotic cells after treatment of cells with P/LOEt as compared with P/LLL. (B) Inhibition of laminin-411 α4 and β1 chain synthesis in human U87MG and T98G glioma cells after treatment with PBS, AON, P/AON/Hu (lacking endosomal escape unit), P/LOEt/AON/Hu, and P/LLL/AON/Hu (1.4 μM with regard to AON). Samples were prepared from the culture supernatant at the end of the treatment and subjected to Western blot analysis. P/LLL/AON/Hu was the most effective in inhibiting the synthesis of both laminin-411 α4 and β1 chains. Secreted fibronectin was used to normalize gel loading.