A kit

RNAs were isolated using TRIzol (Invitrogen), and cDNA was synthesized with a kit (Takara). qPCR was done with SYBR-Green Mix (ABI). The expression change was calculated by 2^−ΔΔCq [15 (link)]. Primers are shown as follows:

Briefly, cells were placed into Trizol (Takara), and RNA was extracted and reverse transcribed into cDNA using a kit (Takara). Relative expression levels of RNA transcripts were determined using gene-specific primers, SYBR green, and a PCR instrument (Bio-Rad).
The sequences of the primers used were as follows: mouse NLRP3 sense 5′-CCAGACCTCCAAGACCACTACG-3′ and antisense 5′-GCTTCCGCAGATCACACTCCT-3′; mouse XO sense 5′-TATGGGGTGGCTTGCTCAGA-3′ and antisense 5′-CAAGACCCTGGACAAATGCC-3′; mouse GAPDH sense 5′-GACATCAAGAAGGTGGTGAAGC-3′ and antisense 5′-GAAGGTGGAAGAGTGGGAGTT-3′.

A kit (TaKaRa) was used for RT-PCR. Total RNAs were isolated with Trizol reagent. RNA (2 μg) was used in the reverse transcription and followed by PCR procedures. The PCR products were examined with agarose gel electrophoresis. GAPDH was used as the internal control in RT-PCR. The sequence of upstream primer for Beclin1: 5′-AGGAGAGACCCAGGAGGAAG-3′; the sequence of downstream primer for Beclin1: 5′-GGCACT TTCTGTGGACATCA-3′; the sequence of upstream primer for GAPDH: 5′-CAGCCAGGAGAAATCAAACAG-3′; the sequence of downstream primer for GAPDH: 5′-GACT GAGTACCTGAACCGGC-3′.