Quantitative Analysis of Gene Expression

Gene expression levels were analyzed using the CFX Connect Real-time PCR (RT-PCR) Detection System (Bio-Rad, Hercules, CA, United States). RT-PCR was performed in duplicate using SsoAdvanced Universal SYBR Green Supermix (Bio-Rad, catalog no. 1725274). The data were analyzed using Bio-Rad CFX Maestro 1.0 software. The specificity of the reactions was determined based on dissociation curve analysis. Fold changes were calculated using the ΔΔCq method as described before (Majewski et al., 2019 (link)). Expression levels were compared between groups using analysis of variance (ANOVA) followed by Tukey’s Honestly Significant Difference (HSD) post hoc test. The 18S ribosomal gene was used as a reference. The sets of primers that were used in the analysis are shown in Table 1.