Journal: mBio

Article Title: MicroRNA-146b protects kidney injury during urinary tract infections by modulating macrophage polarization

doi: 10.1128/mbio.02094-23

Figure Lengend Snippet: miR-146b deficiency promoted M1 macrophage differentiation. (A–D) BMDMs from WT or miR-146b −/− mice were stimulated with CFT073 [multiplicity of infection (MOI) = 0.01] for 6 h. ( A ) Representative flow dot plots and MFI of CD86 in macrophages. Quantification of macrophages, M1 macrophages, M1 macrophages in macrophages, and MFI of CD86 in macrophages. (B–D) Quantitative reverse transcription PCR analysis of the mRNA levels, ELISA, and WB analysis of the protein levels of inflammatory cytokines in infected BMDMs after 6 h. (E–G) THP-1 was treated with 100 ng/mL phorbol 12-myristate 13-acetate for 48 h and then transfected with miR-146b inhibitor. After 48 h, the cells were infected with CFT073 (MOI = 0.01) for 6 h. ( E ) Quantification of macrophages, M1 macrophages, M1 macrophages in macrophages, and MFI of CD86 in macrophages of differentiated THP-1. ( F and G ) ELISA and WB analysis of the levels of inflammatory cytokines in infected THP-1 after 6 h. The data were expressed as the mean ± SEM. The t -test was used to compare between the groups (A–C and E–F). * P < 0.05; ** P < 0.01; *** P < 0.001. All experimental data were collected in at least three independent experiments.

Article Snippet: The cells were then cultured in the 1640 medium with 10% FCS and human GM-CSF (10 ng/mL; Peprotech) for 6 d. THP-1 was cultured in the 1640 medium with 10% FCS and phorbol 12-myristate 13-acetate (100 ng/mL; Sigma-Aldrich) for 48 h. Finally, we obtained BMDMs, HMDMs, and macrophages from THP-1.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Transfection