Fig. 3

NORAD sponges miR-224-3p in ESCC. a The differential miRNAs between CDDP-resistant and CDDP-sensitive ESCC cells in public GSE83362 dataset. b Predicted binding sites on NORAD to sponge miR-224-3p. c The effect of sh-NORAD on the expressions of miR-224-3p, miR-28-5p and miR-7-5p in ESCC cells, normalized to U6 expression. d The expression of NORAD and miR-224-3p in multiple clones of KYSE30/CDDP-R and TE1/CDDP-R cells. NORAD and miR-224-3p was negatively correlated. e NORAD is negatively correlated with miR-224-3p in ESCC specimens while not with miR-28-5p and miR-7-5p. f The expression of miR-224-3p in CDDP-resistant and matched parental ESCC cells, normalized to U6 expression. g The expression of NORAD and miR-224-3p in multiple clones of KYSE30 and TE1 cells. NORAD and miR-224-3p was negatively correlated. h Representative images of RNA FISH of NORAD and miR-224-3p in KYSE30/CDDP-R cells (× 1000), which show that NORAD and miR-224-3p are co-located in the cytoplasm. Nuclei are stained with DAPI. i The nuclear and cytoplasmic expression of NORAD and miR-224-3p in KYSE30/CDDP-R cells. j Luciferase reporter assay showing the luciferase activity of NORAD-WT, NORAD-Mut#1 and NORAD-Mut#2 in KYSE30/CDDP-R and TE1/CDDP-R cells which are co-transfected with miR-224-3p mimic. k Ago2-RIP assay shows that NORAD and miR-224-3p occupy the same Ago2 protein. In all cases, error bars denote SD of triplicates. **P < 0.01, ***P < 0.001