KO1
Peritoneal macrophages (PMs) were isolated from male GSDME-/- and WT mice (n=3) and then treated with ox-LDL (100 g/mL) for 24 hours. After treatment, total RNAs were isolated from macrophages with Trizol reagent (Invitrogen,?Carlsbad, CA, USA) and the sequencing libraries were prepared. Poly-A-containing mRNA was isolated from the total RNA by?poly-T?oligo-attached magnetic beads, and then fragmented by RNA fragmentation kit. The cDNA was synthesized using random primers through?reverse transcription. After the ligation with adaptor, the cDNA was amplified by 15 cycles of PCR, and then 200-bp fragments were isolated using?gel electrophoresis.?The products were sequenced by an Illumina HiSeq2500 instrument in Shanghai Majorbio Biopharm Technology Co. Ltd. (Shanghai, China).