Fig. 6

LINC00680 acts as a miRNA sponge for miR-423-5p to regulate the expression of PAK6 and the malignant behaviors in ESCC cells. a-b Sequence match between miR-423-5p and wild-type (WT) LINC00680 (a) or the 3’ UTR of PAK6 (b) as well as the corresponding mutant form (MT) with the predicted miR-423-5p binding site mutated is shown. c-d KYSE510 (c) and KYSE140 (d) cells were transfected with luciferase reporter vectors containing wild-type (WT-luc) or mutated (MT-luc) LINC00680 in the presence or absence of negative control miRNA (miR-NC) or miR-423-5p mimic (miR-423-5p) followed by dual-luciferase reporter assay. e–f KYSE510 (e) and KYSE140 (f) cells were transfected with luciferase reporter vectors containing wild-type (WT-luc) or mutated (MT-luc) 3’ UTR of PAK6 in the presence or absence of negative control miRNA (miR-NC) or miR-423-5p mimic (miR-423-5p) followed by dual-luciferase reporter assay. g, h, i, j, k, l, n, p, r, t, v KYSE510 and KYSE140 cells were transfected with negative control siRNA (si NC) or siRNA targeting LINC00680 (si LINC00680) in the presence or absence of miR-423-5p inhibitor followed by RT-qPCR analysis (g, h), immunoblotting analysis (i), cell proliferation assay (j, k), colony formation assay (l, n), wound healing assay (p, r), and transwell assay (t, v). Scale bar: 50 µm. m, o, q, s, u, w Quantification of the number of colonies (m, o), the percentage of wound area (q, s), and the number of invasive cells (u, w). The experiments were repeated for three times, and representative data is shown with mean ± standard deviation (SD), *P < 0.05, **P < 0.01, ***P < 0.001