The relative mRNA expression of miR-1224-5p, NLRP3, ASC, FOXO1 and TXNIP was quantified by real-time PCR [29 (link),30 (link)]. Total RNA was obtained from the ovarian tissue of mice or KGN cells by an RNA extraction kit (Tiangen Biotech, Beijing, China). The concentration was quantified by a UV spectrophotometer (Thermo Scientific, Rockford, IL, USA). The RNA sample was then reversibly transcribed into cDNA in a PCR system (Bioneer Corporation, Daejeon, Korea). Quantitative real-time analysis was conducted in the PCR system using the cDNA, SYBR Green (Solarbio Science & Technology), primers (Genscript, Nanjing, China) and 2× Taq PCR Master Mix (Tiangen Biotech). Data were analyzed by 2−ΔΔCt formula and normalized to U6 or GAPDH. The sequences of primers are shown in Table 1.

The sequences of primers

GeneForward primer (5ʹ-3ʹ)Reverse primer (5ʹ-3ʹ)
Mus musculus-miR-1224-5pGTGAGGACTGGGGAGGTGGTGCAGGGTCCGAGGTATTC
Homo sapiens-miR-1224-5pGTGAGGACTCGGGAGGTGGTGCAGGGTCCGAGGTATTC
NLRP3TTCGGAGATTGTGGTTGGGAGGGCGTTGTCACTCAGGT
ASCTGGATGCTCTGTACGGGAAGGTTGTTGCTGGGAAGGAGCCTC
FOXO1GTCCTACGCCGACCTCATCTTGCTGTCACCCTTATCCTTG
TXNIPGTCATCAGTCAGAGGCAATCAAGGAACGCTAACATAGATCAGTAA
Free full text: Click here