Fig. 8

STAT6 binds miR-34a promoter to activate its transcription. a STAT6 overexpression or silencing conducted in M2 macrophages by transfection of STAT6-overexpressing or si-STAT6 vector into M0 and then polarizing to M2, as confirmed by Immunoblotting. b The expression of miR-34a in STAT6-overexpressing or STAT6-silenced M2 macrophages determined by real-time PCR. c A schematic diagram showing the predicted binding sites between STAT6 and miR-34a promoter. Wild- and mutant-type miR-34a luciferase reporter vectors are constructed. d STAT6 and wt- or mut-miR-34a were co-transfected in M0 macrophages followed M2 polarization; the luciferase activity was determined. e The real-time ChIP assay showed that the level of STAT6 antibody binding to miR-34a promoter was much greater than that of IgG. f In macrophages, lncGBP9 competed with SOCS3 for miR-34a binding to counteract miR-34a-mediated suppression on SOCS3, therefore modulating STAT1/STAT6 signaling and the polarization of macrophages. STAT6 bound the promoter of miR-34a to activate its transcription, therefore forming two different regulatory loops to modulate the polarization of macrophages after SCI. Values are mean ± S.D of n = 3 independent experiments