The in vivo antitumor effects of SPION@Au-CS-DOX-FA in a mice tumor model. Changes in the tumor size in representational mice (four groups). (a,b) on day 19 the tumor growth in mice treated with SPION@Au-CS-DOX-FA was reduced significantly (p < 0.01) relative to that of the other groups received free DOX, SPION@Au-CS-FA NPs, and PBS. (c) the proliferation index of splenocytes following the induction of tumor lysate, PHA (positive control), and medium (negative control). (d) splenocytes collected from mice treated by SPION@Au-CS-DOX-FA were re-stimulated with tumor lysate. A substantial increase (p < 0.05) in IFN-ÿ levels compared with those of other groups and a significant decrease (p < 0.05) in IL-4 production was observed in the mice receiving SPION@Au-CS-DOX-FA nanoformulation compared with those given DOX, SPION@Au-CS-FA, and PBS. Results indicate the mean of measurements carried out in triplicate (mean ± S.D., (* p < 0.05; ** p < 0.01; *** p < 0.001); n = 3).