Fitc mouse anti rabbit igg

RAW264.7 cells were transfected with miR-19a-3p mimics, miR-19a-3p mimics NC, miR-19a-3p inhibitor, and miR-19a-3p iNC for 24 h respectively. After that, the cells were stimulated with LPS and IFN-γ for 24 h. Fixed by 4% paraformaldehyde for 30 min, the cells were infiltrated with 1% Triton X-100 for 10 min, and blocked in 10% normal mice serum for 30 min. After incubation with STAT1 (Cell Signaling Technology) or IRF1 antibody (Cell Signaling Technology) overnight at 4°C, FITC Mouse Anti-Rabbit IgG (BOSTER, Wuhan, China) was incubated for 1 h. The nuclei were counterstained using DAPI, and the sections were observed under fluorescence microscope (IX73, Olympus, Japan).

RAW264.7 cells were plated in 24-well culture plates at a concentration of 5×10⁴ cells/well and transfected with miR-103 mimics or NC. After 24 h, the cells were stimulated by IFNγ and LPS for 24h. Thereafter, the treated cells were fixed with 4% paraformaldehyde at room temperature for 30 minutes and then incubated in 10% normal mice serum for 30 minutes. The cells were then incubated with STAT1 antibody (Cell Signaling Technology, Danvers, MA, USA) or IRF1 (8478, Cell Signaling Technology, Danvers, MA, USA) overnight at 4°C. The secondary antibodies FITC Mouse Anti-Rabbit IgG (BOSTER, Wuhan, China) was used for 1 hour. Sections were counterstained with DAPI and analyzed with a fluorescence microscope (IX73, Olympus, Japan).