Figure 4.

(a) Effect of siKRAS entrapped in EPM on the growth of subcutaneous lung tumors growth (a1), weight (a2), and modulation of the target gene in the tumor tissues (a3) in female nude mice. Animals were subcutaneously inoculated with human A549 cells (2.5 × 10⁶ cells) and when tumors grew to 80–100 mm³, animals were randomized (n = 8–10 each) and treated intravenously three times a week (Monday, Wednesday, and Friday) with siKRAS or siSCR (scrambled of siKRAS) entrapped in folic acid (FA)-functionalized EPM. Tumor growth was measured with a caliper weekly; at the end of the study, tumors were weighed and analyzed for the expression of the target protein by western blot. (b) Effect of siKRAS entrapped in EPM on the growth of orthotopic lung tumors (b1) and modulation of the target gene in the tumor tissues (b2) in female NOD Scid mice. Animals were inoculated with Bioware® Brite A549 Red-FLuc lung cancer cells (2 × 10⁶ cells) in 50 μl Matrigel via intrathoracic injection. Ten days later, animals were imaged, randomized, and treated with siKRAS entrapped in EPM, with and without FA-functionalization of exosomes. Two additional groups received FA-EPM or vehicle as controls (n=10 each group). Animals were given luciferin substrate (80 mg/kg, i.p.), 15 min before imaging. Tumor growth was measured in live animals using AMI1000 Imager; at the termination of the study, tumors from randomly selected animals (n=3) were analyzed for the expression of the target protein; data represent mean ± SD. FC, fold change. Statistical analysis was performed by student’s t-test; *, p < 0.05; **, p < 0.01; *** p < 0.001.