Fig 1: Exosomal hsa_circ_0046060 of hUMSC regulates glucose homeostasis and induces insulin resistance through hsa-miR-338-3p in vitro and in vivo. (a) mmu-miR-338-3p expression level (∗∗∗∗P < 0.0001, vs. Control; ^ ^ ^ ^ P < 0.0001, vs. GDM; ####P < 0.0001, vs. GDM + G-mUMSC-EXOs; &&&&P < 0.0001, GDM + C-mUMSC-EXOs vs. GDM + si-circ-G-mUMSC-EXOs) and (b) G6PC2 mRNA expression level (∗P < 0.05, ∗∗∗∗P < 0.0001, vs. Control; ^ P < 0.05, GDM vs. GDM + si-circ-G-hUMSC-EXOs; ##P < 0.01, ###P < 0.001, vs. GDM + G-mUMSC-EXOs; &&&&P < 0.0001, GDM + C-mUMSC-EXOs vs. GDM + si-circ-G-hUMSC-EXOs) in GDM mice administered with PBS, C-mUMSC-EXOs, G-mUMSC-EXOs and si-circ-G-mUMSC-EXOs as evaluated by RT-qPCR. U6 was used as an internal control. (c, d) Protein expression levels of G6PC2 in GDM mice administrated with PBS, C-mUMSC-EXOs, G-mUMSC-EXOs and si-circ-G-mUMSC-EXOs as determined by western blot. GAPDH was used as an internal control (∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001, vs. Control; ^ ^ ^ P < 0.001, ^ ^ ^ ^ P < 0.0001, vs. GDM; ####P < 0.0001, vs. GDM + G-mUMSC-EXOs; &P < 0.05, GDM + C-mUMSC-EXOs vs. GDM + si-circ-G-mUMSC-EXOs). Expression levels of (e) hsa-miR-338-3p (∗∗∗∗P < 0.0001, vs. si-circ-G-hUMSC-EXOs; ^ ^ ^ ^ P < 0.0001, vs. si-circ-G-hUMSC-EXOs + inhibitor NC), (f) G6PC2 (∗∗∗∗P < 0.0001, vs. si-circ-G-hUMSC-EXOs; ^ ^ ^ ^ P < 0.0001, vs. si-circ-G-hUMSC-EXOs + inhibitor NC) and (g) hsa_circ_0046060 (∗∗P < 0.01, vs. si-circ-G-hUMSC-EXOs; ^ ^ P < 0.01, vs. si-circ-G-hUMSC-EXOs + inhibitor NC) following treatment with inhibitor NC and hsa-miR-338-3p inhibitor of si-circ-G-hUMSC-EXOs in L-02 cells as determined by RT-qPCR assay. Glucose concentration: (h) in si-circ-G-hUMSC-EXOs group alone and with inhibitor NC and hsa-miR-338-3p inhibitor (∗∗∗∗P < 0.0001, vs. si-circ-G-hUMSC-EXOs; ^^^^P < 0.0001, vs. si-circ-G-hUMSC-EXOs + inhibitor NC), and (i) PA-induced insulin resistance cell model alone and with inhibitor NC and hsa-miR-338-3p inhibitor of L-02 cells (PI : PA + insulin, ∗∗∗P < 0.001, vs. PI + si-circ-G-hUMSC-EXOs; ^ ^ ^P < 0.001, vs. PI + si-circ-G-hUMSC-EXOs + inhibitor NC) as determined using glucose content assay kit. (j) Glycogen levels in insulin resistance model of L-02 cells as determined using Glycogen Content Assay Kit (PI : PA + insulin, ∗∗P < 0.01, vs. PI + si-circ-G-hUMSC-EXOs; ^ ^ P < 0.01, vs. PI + si-circ-G-hUMSC-EXOs + inhibitor NC). (k) Protein expression levels of (l) G6PC2 (PI : PA + insulin, ∗∗P < 0.01, vs. PI + si-circ-G-hUMSC-EXOs; ^ ^ P < 0.01, vs. PI + si-circ-G-hUMSC-EXOs + inhibitor NC), (m) p-IRS-1 and IRS-1 (PI : PA + insulin, ∗∗P < 0.01, vs. PI + si-circ-G-hUMSC-EXOs; ^ ^P < 0.01, vs. PI + si-circ-G-hUMSC-EXOs + inhibitor NC) in PA-induced insulin resistance cell model alone and with inhibitor NC and hsa-miR-338-3p inhibitor of in L-02 cells as determined by western blot analysis. (n) Morphological changes of liver from GDM mice as evaluated by HE staining. Scale bar = 100 μm.