DHMBA BHT CGA Catechin VC VE DHA EPA 0 62.5 125 250 500 Concentration (mM) Supplemental Figure 1. Representative phase-contrast micrographs of C3A cells treated with DHMBA, BHT, chlorogenic acid (CGA), catechin, vitamin C (VC), vitamin E (VE), docosahexaenoic acid (DHA), and eicosapentaenoic acid (EPA). C3A cells were seeded onto a 24-well plate and incubated with the antioxidants and polyunsaturated fatty acids at concentrations of 0-500 mM for 24 h.
Supplemental Figure 2. Representative flow cytometric quadrantal diagrams on DHMBA and CGA. Apoptotic and necrotic cells (C3A) are plotted in the areas of R3 and R5, respectively. X-axis, propidium iodide; Y-axis, Annexin V. 0 μM 125 μM 250 μM 500 μM CGA DHMBA 22.6% 22.2% 7.23% 8.42% 8.09% 7.52% 6.75% 2.30% 4.49% 5.86% 24.9% 18.8% 12.7% 6.30% 24.1%
5 10 Time (min) 20 40 60 80 100 Relative Abundance 1.3 A 1.7 C [M-H]- 153.00 153.05 m/z 50 153.0196 C7H5O4 B 166 168 169.0509 C8H9O4 D Supplemental Figure 3. Qualitative profiles of oyster extracts obtained by use of LC/LTQ Orbitrap in negative-ion mode. A and B show the extracted ion mass chromatogram and mass spectrum, respectively, of gentisic acid. In the same way, corresponding data for DHMBA, daidzein, and matairesinol are shown in C/D, E/F, and G/H, respectively. 1.6 E 253.0 253.0494 C15H9O4 F 2.3 G 355 360 357.1345 C20H21O6 H
Supplemental Table 1 Phenolic compounds identified in oyster extracts from spectra by LC–LTQ Orbitrap in negtive-ion mode. Theo. Mass Exp. Mass Mass accuracy Phenolic Element [M-H]- Compounds Composition (m/z) (ppm) Gentisic Acid C7H5O4 153.0193 153.0196 1.96 DHMBA C8H9O4 169.0506 169.0509 1.77 Daidzein C15H9O4 253.0506 253.0494 -4.74 Matairesinol C20H21O6 357.1344 357.1345 0.28 Theo. Mass, theoretical mass; Exp. Mass, experimental mass.
Supplemental Table 2. Polyphenols detected and not detected in oyster extracts. Classification Polyphenol Detection non-flavonoids caffeic acid nd DHMBA detected dithranol ferulic acid gallic acid gentisic acid hydroxytyrosol matairesinol phloridzin resveratrol secoisolariciresinol flavonoids apigenin catechin cyanidin epigallocatechin eriodictyol daidzein genistein kaempferol luteolin malvidin naringenin quercetin taxifolin nd, not detected.