Fig. 2

TUSC7 regulated the expression of miR-224. a Online bioinformatics software predicted there were two potential binding sites between TUSC7 and miR-224, and we mutated the two sites. b Luciferase reporter gene vector containing TUSC7 WT or TUSC7 mutant 1 or (and) TUSC7 mutant 2, and miR-224 or pre-NC were co-transfected into HEK293T cells. Dual-luciferase reporter gene assay showed that miR-224 mimic decreased the activity of TUSC7 WT, TUSC7 mutant 1 and TUSC7 mutant 2, and there was no significant difference in the activity in TUSC7 mutant 1 + mutant 2, suggesting miR-224 could bind to the two sites of TUSC7. c miR-224 expression was upregulated in ESCC tissue. d-e EC9706 or KYSE30 cells were transfected with si-NC or si-TUSC7-1 or si-TUSC7-2. qRT-PCR showed that si-TUSC7-1 or si-TUSC7-2 downregulated TUSC7 expression (d), and increased miR-224 expression (e). f-g EC9706 or KYSE30 cells were transfected with pcDNA or pcDNA-TUSC7. qRT-PCR showed that pcDNA-TUSC7 upregulated TUSC7 expression (f), and decreased miR-224 expression (g). h. KYSE30 cell lysate was treated with Ago2 antibody for RNA immunoprecipitation (RIP). qRT-PCR showed that TUSC7 was significantly increased in Ago2 than IgG. i. TUSC7 expression and miR-224 expression was negatively correlated in ESCC tissues