Mir 155 5p inhibitor

M2 macrophages were transfected with plasmids expressing inhibitor NC (NC of miR-155-5p inhibitor) or miR-155-5p inhibitor.
Meanwhile, SW48 cells were transfected with plasmids expressing mimic NC (NC of miR-155-5p mimic), miR-155-5p mimic, inhibitor NC, miR-155-5p inhibitor, sh-NC (scramble shRNA control), sh-ZC3H12B, oe-NC (empty virus vector), oe-ZC3H12B, oe-IL-6, oe-ZC3H12B + oe-IL-6, miR-155-5p mimic + oe-NC, or miR-155-5p mimic + oe-ZC3H12B.
The miR-155-5p mimic and miR-155-5p inhibitor and their controls were purchased from Guangzhou RiboBio (Guangdong, P.R. China). Upon reaching 85–90% cell confluence, M2 macrophages or SW48 colon cancer cells were transfected with plasmids (80 nM) according to the manuals for Lipofectamine 2000 reagents (Invitrogen, Carlsbad, CA, USA). pGCSIL-PUR lentivirus encoding human ZC3H12B (sh-ZC3H12B) and Lenti-OE lentivirus overexpressing IL-6 or ZC3H12B were purchased from Shanghai Genechem (Shanghai, P.R. China). After 6 h of transfection the medium was renewed, and the cells were collected for subsequent experiments after 48 h.
Cy3-labeled miR-155-5p was transfected into M2 macrophages with the Lipofectamine 2000 reagent (Invitrogen). Macrophages containing Cy3-miR-155-5p were cocultured with SW48 cells and observed and detected with a fluorescence microscope and flow cytometry.

KGN cells were a generous gift from the Institute of Applied Anatomy and Reproductive Medicine, Hengyang Medical College, University of South China. The KGN cells were cultured with high glucose Dulbecco's modified Eagle’s medium (DMEM, #11965084, Gibco, Waltham, USA) with 10% foetal bovine serum (FBS, #10099141C, Gibco) in a humidified atmosphere with 5% CO₂ at 37 °C. For the in vitro cellular model of PCOS, KGN cells were cultured with high glucose DMEM with 10% foetal bovine serum (FBS) and 100 nM testosterone (#M6105, AbMole BioScience, Houston, USA) in a humidified atmosphere with 5% CO₂ at 37 °C.
The miR-143-3p mimics, miR-155-5p mimics, miR-143-3p inhibitor and miR-155-5p inhibitor were purchased from RiboBio (Guangzhou, China). Cell transfection was performed using the riboFECT CP Transfection Kit (#C10511-05, RiboBio) when the cells reached 50–60% confluence. The transfection efficiency was evaluated by a riboMONITOR Transfection Indicator Kit (#C10410-5, RiboBio) at 12 h after transfection.