Figure 8.

Magnolol did not increase tumor progression in vitro and in vivo. Effect of magnolol on tumor cell proliferation was determined using the MTS assay in vitro. (A) CT-26 and (B) LLC tumor cell proliferation after magnolol treatment at different concentrations (0, 0.1, 1, and 10 μM). (C) For in vivo assay, LLC tumor-bearing mice were administered vehicle (con), magnolol (mag), cisplatin (cis), or both (cis+mag). The graph shows mean tumor volume (mm³) measured using caliper. ***P < 0.001 vs. con and ^##P < 0.01; ^###P < 0.001 vs. mag based on the two-way Bonferroni post-test at a different time point. (D–H) Tumor tissues were dissociated into single cells and tumor-infltrated immune cells were analyzed by flow cytometry. (D) Gating strategy for analysis of tumor-infiltrated macrophages. Red dots show isotype controls and black dots indicate the stained cells with specific antibodies. (E) Representative FACS plots of CD11b vs. F4/80 and (F) bar graph showing the percentages of CD11b⁺F4/80⁺ macrophages in CD45⁺ cells. (G) Representative dot plots of CD163⁺CD206⁺ M2c macrophages gated on CD11b⁺F4/80⁺ cells and (H) mean percentages of M2c macrophages in tumor tissue. All graphs are expressed as the mean ± SEM of 5 mice. **P < 0.01 vs. con based on the one-way ANOVA using Tukey's test. The letters for no significance were not shown.