*3.4. NMR Analysis*

The samples were dissolved in the deuterated solvent acetone-*d*6 and put into a 5 mm NMR tube. All experiments were performed on a Bruker DRX-500 NMR spectrometer (Bruker, Rheinstetten, Germany), operating at a frequency of 500 MHz for 1H NMR observation, and 125 MHz for 13C NMR observation (at room temperature). The 2D NMR experiments included heteronuclear single-quantum correlation (HSQC) and heteronuclear multiple-bond correlation (HMBC). NMR spectra were carefully processed with the TOPSPIN2.1 ®software (Bruker). The spectra recorded in acetone-*d*6 were referenced to the solvent signal at δH 2.05 ppm and δC 29.92 ppm.

#### *3.5. Preparation of BMDC*

The ICR mice used in this study were obtained from the National Laboratory Animal Center (NLAC, Taipei, Taiwan). The mouse bone marrow-derived DCs were prepared as described previously [31]. Bone marrow cells were isolated from tibias and femurs and then seeded on 6-well plates (Corning) in 4 mL/well RPMI 1640 medium (Thermo), with 10% FBS and 10 ng/mL recombinant mouse GM-CSF

and IL-4 (Peprotech). On day 3 and 5, a 2 mL/well fresh medium containing 10 ng/mL GM-CSF and IL-4 was added. On day 7, BMDCs (> 80% CD11c+ cells) were harvested and used for all experiments.

#### *3.6. Measurement of Cytokine Production*

Cytokine production was measured using an enzyme-linked immunosorbent assay (ELISA), as described previously [31]. The DCs were treated with 100 ng/mL lipopolysaccharide (LPS) (Sigma) or LPS + sample for 6 h for TNF-<sup>α</sup>, IL-6, and IL-12p70 determination. The production of cytokines was measured using the ELISA kit (eBioscience).
