Fig. 3

CRKL is a target of miR-335. a MiR-335 target-site residues in the CRKL 3′UTR are highly conserved among different species. b Direct interaction between miR-335 and CRKL was detected by dual-luciferase reporter assay. Over-expression of miR-335 in SGC-7901 cells (SGC-7901/miR-335) significantly decreased the luciferase signal of CRKL/pMIR/WT compared with the negative control (NigmiR), while mutation of the putative miR-335-binding site abolished this suppressive effect. **P < 0.01). c qRT-PCR analysis of miR-335 in SGC-7901 cells transfected with miR-335-mimic or NC miRNA. Relative expression of miR-335 in SGC-7901 cells transfected with miR-335 mimic was significantly higher than in NC cells. ***P < 0.001 between groups. d qRT-PCR analysis of miR-335 in SGC-7901 cells transfected with miR-335 inhibitor or NC. Relative expression of miR-335 in SGC-7901 cells transfected with miR-335 inhibitor was significantly lower than in NC cells. ***P < 0.001 between groups. e Effect of miR-335 on CRKL mRNA levels in SGC-7901 cells detected by qRT-PCR. Over-expressing miR-335 had no significant effect on CRKL mRNA levels in SGC-7901 cells. f Effect of miR-335 on CRKL protein expression in SGC-7901 cells detected by western blot analysis. CRKL protein expression was significantly suppressed by over-expressing miR-335 in SGC-7901 cells