Fig. 4

CRAT16 functions as a miR-371a-5p sponge. a FISH analysis of CRART16 in Caco-2 and Caco-2 CR cells (nuclei were stained with DAPI). Scale bar = 25 μm. b MiRNA heatmap shows the top differentially expressed miRNAs in Caco-2-CRART16 cells versus Caco-2-NC cells. c Schematic diagram of the predicted binding sites between lncRNA CRAT16 and miR-371a-5p. d The expression of miR-371a-5p was detected in Caco-2, Caco-2 CR, Caco-2-CRART16 and Caco-2-NC cells by qRT-PCR. U6 was used as an internal control. All experiments were repeated three times, and data are presented as mean ± SD. *P < 0.05, **P < 0.01 by Student’s t test. e Dual-luciferase reporter assays in 293T cells. The relative luciferase activity was measured after cotransfection with miR-371a-5p mimics or NC and either the pmiR-RB-Report™-CRART16-WT vector or the empty vector. All experiments were repeated three times, and data are presented as mean ± SD. **P < 0.01 by Student’s t test. f The expression of miR-371a-5p was detected by qRT-PCR in Caco-2-CRART16 cells after transfection of miR-371a-5p mimics. U6 was used as an internal control. All experiments were repeated three times, and data are presented as mean ± SD. *P < 0.05, **P < 0.01 by Student’s t test. g CCK8 assay was performed to assess the cell viability of Caco-2-CRART16 cells after miR-371a-5p overexpression with graded concentrations of cetuximab (0–200 μg/ml) for 48 h. Each concentration had six replicates, and the experiment was repeated at least three times. Data are presented as mean ± SD. **P < 0.01 by two-way ANOVA