Fig. 4

Adipose-derived exosomal miR-421 targets CBX7. A Venn diagram analysis from predicted miRNAs targeting CBX7 from mirTarBase (Table 2) and TargetScanHuman (Table 3) and lists of adipocyte specific miRNA from Adicer KO mice and patients with congenital lipodystrophy (CGL) [64] shows miR-421 as the only common element in the overlap; B OCSC1-F2 human ovarian cancer cells were treated with adipose-derived exosomes (exo) in the presence of anti-miR-421 or anti-miR negative control (-ve con Anti-mir) and effect on CBX7 was determined by Western blot analysis; NT, no treatment control (C) OCSC1-F2 human ovarian cancer cells were treated with miR-421 mimic and the effect on CBX7 was determined by Western blot analysis; Neg, non-specific miRNA; D plasmid design for luciferase reporter system carrying CBX7 3’ UTR predicted to be the binding site for miR-421 (pCBX7) and its mutated version (pCBX7mut). Sequence of human miR-421 (hsa-mir-421) is also shown; E Luciferase reporters were transfected in OCSC1-F2 human OC cells in the presence or absence of miR-421 or non-specific miRNA (Neg) as indicated, and levels of luciferase were measured as surrogate of binding affinity; F miR-421 was quantified in adipose- derived exosomes using qPCR. Note that exosomes successful in downregulating CBX7 (columns with red data points) contain higher levels of mir-421. Data are presented as mean ± SEM (n = 3). Ordinary One way ANOVA with Tukey’s multiple comparison test was used to determine statistical significance