Results were calculated using the ΔΔCt method and expressed as a percentage of expression compared with SH-SY5Y cells±S.D., with EFNB2 mRNA in SH-SY5Y cells assigned an expression level of 100%.
The mRNA expression was determined using the Livak method and expressed as fold changes [ 31].
Gene expression levels normalized to endogenous control gene expression were analyzed using the ΔΔCt method and expressed as mean percentages ± SEM relative to the WT condition.
Mean (±SEM) fold change in Glast expression for Clock/Clock astrocytes was calculated using the ΔΔCt (deltadeltaCt) method and expressed as a function of wild-type values.
Relative mRNA expression levels were quantified using the comparative Ct(Δ Ct) method and expressed as 2− ΔΔ C t).
These methods are mode superposition method and expressing governing equations in state space format.
Infrared (IR) spectra was recorded on Bruker 12,060,280, Software: OPUS 7.2.139.1294 spectrophotometer by KBr pellet method and expressed in cm−1.
Antibacterial activity was done by disc diffusion method and expressed as in millimeter diameter of zone inhibition.
Finally, three variant-encoding genes, Aoxyn11A G21I, Aoxyn11A Y13F and Aoxyn11A G21I–Y13F, were constructed by two-stage whole-plasmid PCR method, and expressed in Pichia pastoris GS115, respectively.
Because of the significant matrix effect found, analyses of wine samples towards polyphenol contents were carried out using standard addition method and expressed as gallic acid equivalents.
