Fig. 3

TCTN1 was a direct target of miR-216a-5p. a Putative binding sites of 216a-5p within the 3′-UTR region of TCTN1 mRNA, and the sequences of wild-type and mutant-type vector. b The relative luciferase activities were inhibited in 293 T cells co-transfected with wild-type TCTN1 3′-UTR vector and miR-216a-5p, not with the mutant-type vector. Firefly luciferase activity was normalized to Renilla luciferase. Data are presented as the mean value ± SD from triplicate experiments. ***p < 0.001 vs. miR-NC; c mRNA and d protein levels of TCTN1 were detected by qRT-PCR and western blot in EC9706 and TE-9 cells transfected with miR-216a-5p or miR-NC. e The protein level of TCTN1 was determined by western blot in HET-1A cells transfected with miR-216a-5p inhibitor or miR-NC. f Relative expression levels of TCTN1 mRNA in ESCC tissues and adjacent tissues were detected by qRT-PCR. g Pearson’s correlation analysis for the relationship between miR-216a-5p levels and TCTN1 mRNA levels in ESCC tissues