Fig. 6

Exosome circATP8A1 promotes macrophage M2 polarization by regulating the STAT6 pathway through competitive binding of miR-1-3p. A RIP results showed enrichment of circATP8A1 in Ago2 immunoprecipitations (P < 0.05, n = 3). B RIP results showed enrichment of miR-1-3p in Ago2 immunoprecipitation (P < 0.05, n = 3). C The relative level of circATP8A1 was determined by qPCR after pull-down with a biotin-labeled miR-1-3p probe or oligo probe. D The relative level of miRNAs was determined by qPCR after pull-down with a biotin-labeled circATP8A1 probe or control probe. E Luciferase assay of HEK-293 T with Luc-circATP8A1-wt or Luc-circATP8A1-mut co-transfected with miR-1-3p mimic or mimic normal control. F Luciferase assay of HEK-293 T with Luc-STAT6-wt or Luc-STAT6-mut co-transfected with miR-1-3p mimic (miR1 mimic) or mimic normal control (mimic NC). G-J Rescue experiments were conducted to confirm the interaction between circATP8A1 and miR-1-3p. M0-type macrophages were treated with SGC7901 exosomes overexpressing circATP8A1 and further with miR1-3p mimic and mimic control. qPCR was used to detect the expression of M1 and M2 polarization markers. K Western blot analysis was conducted to examine the expression levels of the STAT6, pSTAT6 and CD206 in the experimental samples. * P < 0.05, ** P < 0.01, *** P < 0.001. NS, not significant