The protein concentration in RIPA extracts was quantitated by Lowry's method [ 78].
The isoflavone content of the extracts was quantitated by high-performance liquid chromatography (HPLC; Waters 2695 system, Waters Co., Milford, MA, USA) using a CAPCELLPAK UG 120 250 × 4.6 mm, 5-µm column (Shiseido Co., Tokyo, Japan).
The total amount of RNA extracted was quantitated by spectrometry at 260 nm, and its purity was evaluated by the ratio between the absorbance values at 260 and 280 nm, whereas its integrity was confirmed in agarose gels.
RNA extracts were quantitated with Quant-iT™ RiboGreen ® RNA Kit (Invitrogen by Life Technologies, Carlsbad, CA) as previously described.
S1P levels in dried extracts were quantitated using a whole cell receptor binding assay [59].
DNA extracts were quantitated using the Quantifiler™ Human DNA Quantification kit (Applied Biosystems, Foster City, CA) according to manufacturer's protocols.
Brains were removed, weighed, homogenized, extracted, and the corresponding phloretin TMS-ether derivatives were quantitated by EI GC MS.
After 2, 6, or 18 hours of infection, cellular DNAs were extracted and viral early or late reverse transcription (RT) products were quantitated by Real-time PCR (Fig. 4D).
Total amounts of protein extracts was quantitated using Bio-Rad protein assay (Pierce, Rockford, IL).
