Fig. 4

PVT1 acted as a miR-23b-3p sponge and WT1 was a direct target of miR-23b-3p. a Schematic of the miR-23b-3p-binding site in PVT1 and WT1 3′-UTR, and mutated miR-23b-3p-binding site. b The luciferase activity in MCs transfected with PVT1 wild-type luciferase reporter plasmid (PVT1-WT), WT1 3′-UTR wild-type reporter construct (WT1-WT), or the site-directed mutant of the target sequence (PVT1-MUT and WT1-MUT) together with miR-NC mimic or miR-23b-3p mimic. c PVT1 expression by qRT-PCR in MCs transfected with PVT1 overexpression plasmid or the negative plasmid. NC: negative plasmid, PVT1: PVT1 overexpression plasmid. d The expression of miR-23b-3p in MCs transfected with si-NC, si-PVT1#1, PVT1 overexpression plasmid, or the negative plasmid. NC: negative plasmid, PVT1: PVT1 overexpression plasmid. MCs were introduced with miR-NC mimic, miR-23b-3p mimic, anti-miR-NC or anti-miR-23b-3p, followed by the determination of miR-23b-3p expression by qRT-PCR (E), WT1 mRNA expression by qRT-PCR (f), WT1 protein level by western blot (g). h MiR-23b-3p expression by qRT-PCR in 34 serum samples from patients with DN and 30 serum samples from healthy controls. *P < 0.05