Fig. 4

HNF1β was upregulated in glioma tissues and cell lines and was a target of miR-217; both circ-TTBK2 and miR-217 could modulate HNF1β expression. a The predicted miR-217 binding site in HNF1β (HNF1β-Wt) or and the designed mutant sequence (HNF1β-Mut) were indicated. b Luciferase reporter assay of HEK 293T cells co-transfected with HNF1β-Wt or HNF1β-Mut and miR-217 or the miR-217-NC (data are presented as the mean + SD (n = 5, each group). ^* P < 0.05 vs. HNF1β-Wt+miR-217-NC group). c Immunohistochemistry of HNF1β protein in nontumorous brain, low-grade glioma, and high-grade glioma tissues. Original magnification: 100×. Scale bar = 50 μm. d HNF1β protein expression levels in nontumorous brain tissues and glioma tissues using GAPDH as an endogenous control. Representative protein expression and their integrated density values (IDVs) of HNF1β in nontumorous brain tissues, low-grade glioma tissues (World Health Organization (WHO) I–II), and high-grade glioma tissues (WHO III-IV) are shown (data are presented as the mean + SD (n = 15, each group). ^** P < 0.01 vs. NBTs group; ^## P < 0.01 vs. low-grade glioma tissues group). HNF1β protein expression levels in astrocytes, U87, and U251 cells and using GAPDH as an endogenous control. Representative protein expression and their IDVs in human normal astrocytes, U87, and U251 are shown (data are presented as the mean + SD (n = 15, each group). ^** P < 0.01 vs. human normal astrocytes group). e qRT-PCR and western blot analysis for circ-TTBK2 regulated HNF1β expression in U87 and U251 cells. The relative expression of HNF1β mRNA was shown using GAPDH as an endogenous control. The IDVs of PIWIL4 was shown using GAPDH as an endogenous control (data are presented as the mean + SD (n = 5, each group). ^* P < 0.05 vs. circ-TTBK2 (+)-NC group; ^# P < 0.05 vs. circ-TTBK2 (−)-NC group). f qRT-PCR and western blot analysis for miR-217 regulated HNF1β expression in U87 and U251. The relative expression of HNF1β mRNA was shown using GAPDH as an endogenous control. The IDVs of HNF1β was shown using GAPDH as an endogenous control (data are presented as the mean + SD (n = 5, each group). ^* P < 0.05 vs. pre-NC group; ^# P < 0.05 vs. anti-NC group). g qRT-PCR and western blot analysis for circ-TTBK2 combined with miR-217 regulated HNF1β expression in U87 and U251. The relative expression of HNF1β mRNA was shown using GAPDH as an endogenous control. The IDVs of HNF1β was shown using GAPDH as an endogenous control (data are presented as the mean + SD (n = 5, each group). ^* P < 0.05 vs. circ-TTBK2 (+)+miR-217 (+) group; ^# P < 0.05 vs. circ-TTBK2 (−)+miR-217 (−) group)