Fig 1: Circ_0000745 upregulates ERBB4 to mediate the PI3K/AKT signaling pathway in OC cells. A, transfection efficacy of sh-ERBB4 and oe-ERBB4 in ES-2 or SK-OV-3 cells examined by RT-qPCR; B, phosphorylation of AKT and PI3K in ES-2 and SK-OV-3 examined by western blot analysis; C, proliferation ability of ES-2 and SK-OV-3 cells examined by the CCK-8 assay; D, colony formation ability of ES-2 and SK-OV-3 cells measured by the colony formation assay; E, migration ability of ES-2 and SK-OV-3 cells examined by the wound-healing assay; F, invasion ability of ES-2 and SK-OV-3 cells determined by the Transwell assay; G, stem cell property of ES-2 and SK-OV-3 cells determined by the sphere formation assay. Data were exhibited as mean ± SD from three repetitions. Differences were analyzed by one-way ANOVA (A, D, E, F and G) or two-way ANOVA (B and C); *p < 0.05 vs. oe-circ + sh-NC; #p < 0.05 vs. sh-circ + oe-NC
Fig 2: miR-3187-3p directly targets ERBB4. A, a Venn diagram for the intersections of the candidate target mRNAs of miR-3187-3p from five bioinformatic systems; B, a KEGG pathway enrichment analysis based on the candidate mRNAs; C, expression of ERBB4 in OC predicted on the GEPIA database; D, mRNA expression of ERBB4 in the collected tissue samples detected by RT-qPCR; E, a negative correlation between the expression of miR-3187-3p and ERBB4 mRNA in OC tissues; F, mRNA expression of ERBB4 in ES-2 and SK-OV-3 cells after oe-circ/sh-circ/miR-mimic/miR-inhibitor transfection examined by RT-qPCR; G, putative binding sequence between miR-3187-3p and ERBB4 3’UTR and the MT sequence for luciferase assay; H, luciferase activity of circ-WT and circ-MT in 293 T cells examined by the luciferase reporter gene assay. Data were exhibited as mean ± SD from three repetitions. Differences were analyzed by paired t test (D), one-way ANOVA (F) or two-way ANOVA (H); *p < 0.05 vs. adjacent; #p < 0.05 vs. oe-NC; &p < 0.05 vs. oe-circ +NC mimic; @p < 0.05 vs. sh-NC; ^p < 0.05 vs. sh-circ + NC inhibitor; %p < 0.05 vs. NC mimic; In panel E, the correlation between miR-3187-3p and ERBB4 was validated using Pearson’s correlation analysis, r = - 0.702, p < 0.001
Fig 3: Silencing of IGF2BP2 suppresses the tumorigenesis of OC cells in vivo. A, expression of circ_0000745 in SK-OV-3 cells examined by RT-qPCR; B, growth rate of xenograft tumors; C, weight of the xenograft tumors on the 35th d; D, expression of IGF2BP2 mRNA, circ_0000745, miR-3187-3p, and ERBB4 mRNA in tumor tissues examined by RT-qPCR; E, protein levels of IGF2BP2 and ERBB4 tumor tissues determined by western blot analysis. In each group, n = 5; the mean value was calculated. Data were exhibited as mean ± SD. Differences were analyzed by the two-way ANOVA (A and C) or two-way ANOVA (B, D and E); *p < 0.05 vs. sh-NC; #p < 0.05 vs. sh-IGF2BP2 + oe-NC
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