Figure3.

Upregulation of IGF2BP2 increased FBXL19-AS1 stability, FBXL19-AS1 participated in the process of IGF2BP2 regulating BTB permeability

(A). Knockdown of IGF2BP2 decreased the expression level of FBXL19-AS1. Data represent mean ± SD (n = 3, each). **P < 0.01 vs. IGF2BP2 (+)-NC group, ^#P < 0.05 vs. IGF2BP2 (-)-NC group. (B) RIP assay evaluated the remarkable enrichment of FBXL19-AS1 in anti-IGF2BP2 immunoprecipitates. Results were analysed using two-tailed paired Student’s t test. Data represent mean ± SD (n = 3, each). **P < 0.01 vs. Anti-normal IgG group. (C) IGF2BP2 and GAPDH protein in immunoprecipitation with FBXL19 and FBXL19-AS1 RNA were visualized by western blot. (D) No significant difference of nascent FBXL19-AS1 was found in silenced IGF2BP2 GECs. (E) Knockdown of IGF2BP2 decreased the half-life time of FBXL19-AS1. Results were analysed using two-tailed paired Student’s t test. Data represent mean ± SD (n = 3, each). **P < 0.01 vs. IGF2BP2 (-)-NC group. (F,G) Overexpression of FBXL19-AS1 significantly rescued the inhibitory or increasing effects of IGF2BP2 knockdown on TEER value and HRP exudate respectively. (H) Overexpression of FBXL19-AS1 significantly rescued the inhibitory effect of IGF2BP2 knockdown on the expression levels of ZO-1, occludin and claudin-5. (I,J) Overexpression of FBXL19-AS1 significantly rescued the increasing effect of IGF2BP2 knockdown 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, ^##P < 0.01 vs. IGF2BP2(-)+FBXL19-AS1(+) group, ^&&P < 0.01 vs. IGF2BP2 (+)+FBXL19-AS1(-) group.