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. 2021 Apr 12;9:640402. doi: 10.3389/fcell.2021.640402

FIGURE 4.

FIGURE 4

CircGFRα1 functions as a ceRNA to sequester miR-449 and upregulate the expression of GFRα1. (A) qRT–PCR analysis for the expression of circGFRα1 after GDNF removal and replenishment. (B) Target region of the 3′-UTR GFRα1 for miR-449. (C) Effects of miR-449 on the activity of firefly luciferase reporters containing either 3′-UTR GFRα1 or mutant type 3′-UTR GFRα1 were assessed by luciferase reporter gene assays. (D) RIP assays were performed to detect miR-449 and 3′-UTR GFRα1. (E) qRT–PCR and western blotting analysis for the expression of GFRα1 in FGSCs infected with the circGFRα1 overexpression lentivirus control (over-con), circGFRα1 overexpression lentivirus (over). (F) qRT–PCR and western blotting analysis for the expression of GFRα1 in FGSCs infected with the circGFRα1 knockdown lentivirus control (kd-con), circGFRα1 knockdown lentivirus (kd). (G) qRT–PCR and western blotting analysis for the expression of GFRα1 in FGSCs infected with the circGFRα1 knockdown lentivirus control (kd-con), circGFRα1 knockdown lentivirus (kd), and co-transfected miR-449 inhibitor and circGFRα1 knockdown lentivirus (kd + inhibitor). ***P < 0.001. n.s., no significant.