*2.6. Statistical Analysis*

All experiments were carried out in at least three independent replicates (*n* = 3), and all data were expressed as the means ± standard deviation. The statistical significance was determined by Duncan's multiple range test (*p*-values < 0.05). For the RSM analysis, the generated 3D surface was determined from the fitted polynomial equation, and significant coefficients (*p* < 0.01) were used in the model. The variance table was generated from both independent variables and experimental outputs using the Design Expert statistical software (version 11.0; State-Ease Inc., Minneapolis, MN, USA). For the pairwise comparisons during the proteomic analysis, we performed One-Way analysis of variance (One-Way ANOVA) at the protein-level analysis with two multiple testing correction methods including the Bonferroni and the Benjamini–Hochberg FDR corrections using the ProteinPilot™ Software.
