Mir 205 5p mimic

To down-regulate circ-FKBP5, small interfering RNA (siRNA) targeting circ-FKBP5 (si-circ-FKBP5) was constructed in U6/GFP/Neo plasmid. oe-circ-FKBP5, oe-NC, si-circ-FKBP5, si-NC, miR-205-5p mimic, mimic NC, miR-205-5p inhibitor, inhibitor NC, sh-RUNX2, and sh-NC were purchased from Genepharma (Shanghai, China). BMSCs were seeded in 2 mL of osteogenic induction medium and transfected using Lipofectamine2000 (Invitrogen). After 24 h of transfection at 37 °C and 5%CO₂, the previous medium was replaced with a fresh medium and incubated for 48 h [24 (link)].

The mouse lung epithelial (MLE‐12) cell lines were purchased from the BeNa Culture Collection. Cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) containing 10% (v/v) fetal bovine serum (FBS), in a humidified atmosphere of 95% air‐ 5% CO₂ at 37°C. MLE‐12 cells were cultured in 6 well culture plate, and then the cells were transfected with miR‐205‐5p mimic, miR‐205‐5p inhibitor, miR‐431‐5p mimic, miR‐431‐5p inhibitor, NC or inhibitor negative control (INC), with lipofectamine 2000 for 48 h. miR‐205‐5p inhibitor, miR‐431‐5p inhibitor, INC were synthesized by Genepharma. These miRNA mimic (or respective inhibitor) transfected MLE‐12 cells were simultaneously infected with influenza A virus (MLE‐12 Infection model). The cells cultured in complete medium were considered as normal control. The culture plate was cultured in a 37°C, 5% CO₂ incubator. The cells were harvested 48 h after transfection, and Western blot was carried out using the primary antibodies against NP (Abcam) and GAPDH (Cell Signalling) as an internal control.²³