Magnolol

Magnolol, purchased from Sigma-Aldrich (St. Louis, MO, USA), was dissolved with 50% aqueous ethanol (v/v) to prepare for the stock solution. It was mixed with the medium to obtain the different concentrations (0, 12.5, 25, 50, 100 and 200 μM). Antifungal trials of Magnolol were performed as our previous work with slight modifications [31 (link)]. A total of 5 μL aliquot of spore suspension was inoculated on PDA medium with serial concentrations of Magnolol and cultured at 25 °C for 6 days for diameter determination. In addition, liquid culture was carried out using potato dextrose broth (PDB; Becton Dickinson, Franklin Lakes, NJ, USA) at the final concentration of 10² spores/mL with shake flask culture in an incubator at 25 °C, 180 r/min. The minimum inhibitory concentration (MIC) was recognized as the lowest concentration of Magnolol without visible fungal extending. After incubation for 6 d, the culture was filtered with Whatman filter paper. The supernatant was used for the determination of AOH and AME contents, while the mycelium was freeze-dried for the biomass analysis or quickly frozen with liquid nitrogen for the following RNA extraction. Each test was performed in triplicate. The inhibition ratio of the mycelial growth and biomass weight was separately calculated.

Magnolol (purity ≥ 95%) was obtained from Sigma (m3445; Sigma-Aldrich, St. Louis, MO, USA). Magnolol was dissolved in DMSO to prepare 10 mM stock solution. Cisplatin was purchased from Sigma-Aldrich (P4394; St. Louis, MO, USA) and dissolved in saline at 1 mg/mL.

Briefly, 3-(4, 5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), magnolol and dimethyl sulfoxide (DMSO) were all purchased from Sigma-Aldrich (St.Louis, MO, USA). In addition, NF-κB inhibitor 4-N-[2-(4-phenoxypheny) ethyl]quinazoline-4,6-diamine (QNZ), AKT inhibitor LY294002, ERK inhibitor PD98059, P38 inhibitor SB203580 and JNK inhibitor SP600125 were all purchased from Selleckchem (Houston, TX, USA).

Oxacillin, honokiol (≥98%) [3V,5-di-2-propenyl-(1,1V-biphenyl)-2,4V-diol], and magnolol (≥95%) [5V,5-di-2-propenyl-(1,1V-biphenyl)-2,2V-diol] were purchased from Sigma. magnolol and honokiol were dissolved in 100% EtOH.

The HSC-3 and SCC-9 oral cancer cells were seeded onto 24-well plates at a density of 8 × 10⁴ cells overnight and then treated with magnolol (M3445, Sigma-Aldrich, St. Louis, MO, USA) at different concentrations (0, 25, 50, 75, and 100 μM) for 24 h. Subsequently, the cells were incubated with MTT solution for 4 h at 37 °C as previously described [17 (link)].

Magnolol was obtained from Sigma-Aldrich (St. Louis, MO, USA). Alpha-modified minimal essential medium (α-MEM) and fetal bovine serum (FBS) were purchased from Thermo Fisher Scientific Inc. (Rockford, IL, USA). Recombinant IL-1α was obtained from PeproTech (Rocky Hill, NJ, USA). Recombinant M-CSF was kindly provided by Dr. Yongwon Choi (University of Pennsylvania School of Medicine, Philadelphia, PA, USA). Recombinant soluble RANKL was prepared as described previously [19 (link)].