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SS determination was conducted using the photometric method, Method 8006.
Protein determination was conducted using a BCA-based protein assay (Pierce Biotechnology/Thermo Scientific).
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All lipid determinations were conducted using standard methods as previously described [ 13].
The pellet was resuspended in buffer A and protein determinations were conducted using the Bio-Rad protein assay.
Electrode surface area determinations were conducted using a 10 mM solution of Fe CN 63 /4– (1:1 ratio mixed solution), in a range of (−0.1) to (+0.6) V vs Ag/AgCl.
Quantitative determination of E2 was conducted using a linear calibration curve between the range of 0.78 25 μg/mL with a 0.99 correlation coefficient.
The determination of CHX was conducted using polystyrene flat-bottom microplate wells (Sarstedt, Nümbrecht, Germany) in a microplate reader (SpectraMax M2, Molecular Devices, Ismaning/Munich, Germany).
The determination of HMF concentrations was conducted using a high-performance liquid chromatography (HPLC) method based on the method published by the International Honey Commission (IHC, 1999).
Copy number determination for each transcript was conducted using real-time competitive PCR coupled with product resolution via Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry (MassARRAY QGE, Sequenom, San Diego, CA)., as previously described [ 13].
Determination of total tannin content was conducted using a similar method for the determination of TPC described by Makkar et al. [18].
Determination of the Haugh unit was conducted using Egg Analyzer™ (SANOVO, Denmark).
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