Quantitative Reverse Transcription-PCR (qRT-PCR)

One microgram of total RNA was used for cDNA synthesis with random hexanucleotides and SuperScript III reverse transcriptase (Life Technologies, USA). Quantitative PCR was performed using qPCRmix-HS SYBR (Evrogen, Russia) and the Light Cycler 480 real-time PCR system (Roche, Switzerland); cycling conditions were as follows: 95°C for 20 s, 61°C for 20 s, 72°C for 30 s, repeat 40 times; primers are listed in Supplementary Table 1. In the end of amplification, a dissociation curve was plotted to confirm specificity of the product. All real-time experiments were repeated in triplicate. The results were normalized against the 16S rRNA gene. Calculations were performed according to (Ganger et al., 2017) for the relative expression ratio.