Total RNA was extracted from dry cell pellets using the RNAeasy kit (Qiagen), per manufacturer’s instructions. Concentration and purity of ribonucleic acid (RNA) was determined using the Nanodrop. 600 ng RNA was converted to cDNA with the QuantiTect reverse transcription kit (Qiagen) as per manufacturer’s instructions. Quantitative real-time polymerase chain reaction (PCR) reactions were performed using the QuantiTect SYBR Green kit (Qiagen). To summarise, 2 mL cDNA, 10 mL SYBR Green, 7 mL RNAse free dH2O and 1 mL 20 mM forward and reverse primers was added to each well. A mock mixture using 2 mL RNAse free dH2O instead of cDNA was used as a blank. Primers used (Eurofins Genomics). For the PCR reaction activation was at 95°C for 10 minutes. Following this was 40 cycles of denaturing at 94°C for 15 seconds; annealing at 58°C for 30 seconds and extension at 72°C for 30 seconds. Fold-change in gene expression was calculated using the ΔCT method, based on biological reference samples and glyceraldehyde 3-phoshate dehydrogenase (GAPDH) mRNA levels for normalisation. All results obtained were from the evaluation of two technical duplicates of three independent experiments.