Pcdna3.1 rock1

THP-1 and RAW 264.7 cells were plated on 12-well plates at a seeding density of 3×10⁵ cells/well and transfected with miR-502-3p mimic, miR-502-3p inhibitor, pcDNA3.1-ROCK1 (Shanghai GenePharma Co., Ltd.), or their negative controls (NCs), mimic NC, inhibitor NC and pcDNA3.1-NC (50 pg/well) at 37°C for 24 h, using Lipofectamine^® 3000 (Invitrogen; Thermo Fisher Scientific, Inc.). At 24 h following transfection, the transfected macrophages were used for other experimental assays. The sequences are listed in Table I.

Small interfering RNAs (siRNAs) for KCNMB2-AS1 (si-KCNMB2-AS1) and negative control (NC) siRNAs (si-NC) were designed and synthesized by GenePharma Co., Ltd. (Shanghai, China). The si-KCNMB2-AS1#1 sequence was 5ʹ-AACTTTTTATTAGATATCAAAGA-3ʹ; si-KCNMB2-AS1#2 sequence was 5ʹ-ATGAAGATATCTAAAAACAAAGA-3ʹ; si-KCNMB2-AS1#3 sequence was 5ʹ-CTGGATTTCTTGTGAAGAAAACT-3ʹ; and the si-NC sequence was 5ʹ-CACGATAAGACAATGTATTT-3ʹ. An overexpression vector (pcDNA3.1/+) harboring human ρ-associated coiled-coil–containing protein kinase 1 (pcDNA3.1-ROCK1) and empty pcDNA3.1 plasmid were also synthesized by GenePharma Co., Ltd. miR-374a-3p mimic, miR-negative control (miR-NC), miR-374a-3p inhibitor, and NC inhibitor were obtained from RiboBio Co., Ltd. (Guangzhou, China). Cells were seeded into 6-well plates, and Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) was used for oligonucleotide and plasmid transfections.