Figure 5.

FBXL19-AS1 negatively regulated ZNF765 expression through SMD pathway

(A) Dual-luciferase reporter assays were performed to determine the binding sites of FBXL19-AS1 and ZNF765 3ʹ-UTR. Data represent mean ±SD (n = 3, each). *P < 0.05 vs ZNF765 3ʹ-UTR-Wt + FBXL19-AS1 (-)-NC group. (B) Knockdown of FBXL19-AS1 significantly induced the enrichment of FBXL19-AS1 and ZNF765 mRNA with STAU1 respectively. Results were analysed using two-tailed paired Student’s t test respectively. Data represent mean ± SD (n = 3, each). *P < 0.05, **P < 0.01 vs. anti-IgG group, ^#P < 0.05 vs. Control+anti-STAU1 group. (C) Overexpression of FBXL19-AS1, knockdown of STAU1 and knockdown of UPF1 reduced the half-life time of ZNF765 mRNA. Results were analysed using one-way ANOVA test. Data represent mean ± SD (n = 3, each). *P < 0.05 vs. Control group. (D,E) Knockdown of STAU1 rescued the inhibitory effect of overexpression FBXL19-AS1 on the mRNA and protein expression levels of ZNF765. Results were analysed using one-way ANOVA test. Data represent mean ± SD (n = 3, each). **P < 0.01 vs. Control group. (F,G) Knockdown of ZNF765 rescued the inhibitory or increasing effect of FBXL19-AS1 knockdown on TEER values or HRP of GECs respectively. (H) Knockdown of ZNF765 rescued the inhibitory effect of FBXL19-AS1 knockdown on the expression of ZO-1, occludin and claudin-5 in GECs. Results were analysed using one-way ANOVA test. Data represent mean ± SD (n = 3, each). **P < 0.01 vs. Control group, ^##P < 0.01 vs. FBXL19-AS1 (+)+ZNF765 (+) group, ^&&P < 0.01 vs. FBXL19-AS1 (-)+ZNF765 (-) group.