Total RNA was extracted using Trizol reagent (Life Technologies, Carlsbad, CA) according to the manufacturer’s instructions. The concentration and the quality of the extracted RNA were evaluated by using Take3 micro-volume plate and the Synergy HT multi-mode microplate reader (BioTek, Winooski, VT). RNA was assessed by using (A260/280) ratio, and those RNAs which were with ratio absorbance >1.8 were judged as high quality and high integrity RNAs. The RNA was loaded onto agarose gel electrophoresis for visual assessment. The reverse transcription of the RNA (1 μg) was done with qScript® cDNA Synthesis SuperMix (Quanta Biosciences, Qiagen, Beverly Inc.). The resulting cDNA was amplified by using qPCR (Applied Biosystems 7500 Real-Time System) with SYBR green master mix (Life Technologies, Carlsbad, CA) according to the manufacturer’s instructions. Relative quantification (RQ) of the mRNA expression of the NLRP3 gene was determined using the 2-ΔΔCT calculation (Pfaffl, 2004 ; Schmittgen and Livak, 2008 (link)). All readings were normalized to 18S ribosomal RNA expression as the housekeeping gene (Kuchipudi et al., 2012 (link)). Primers designed for mRNA of the NLRP3 and 18S rRNA genes were as described by Kuchipudi et al. (2012) (link) and Karaffová et al. (2020) (Table 1).