Fig. 2: LIMK2 is upregulated in ESCC and promotes ESCC growth in vitro and in vivo.

A Representative images of IHC staining of LIMK2 in ESCC tissue microarray. Magnification ×40 and ×100. Scale bars respectively represents 250 µm and 50 µm. B Expression of LIMK2 in normal and tumor ESCC tissues (Adjacent = 66, Cancer = 107). C Expression of LIMK2 in paired normal and tumor ESCC tissues (n = 65). D Expression level of LIMK2 among ECA and ESCA from TCGA database. E Relationship between LIMK2 expression and the overall survival of patients from TCGA database. F Western blot analysis of LIMK2 expression in SHEE cells and seven human ESCC cell lines. GAPDH served as a control. G Western blot analysis of LIMK2 in knock down cells. GAPDH served as a control. H Indicated cells were subjected to soft agar assays, and the relative colony numbers were quantified (n = 3, independent experiments). I MTT assay showed the proliferation ability of LIMK2-shRNA treated cells (n = 3, independent experiments) J Western blot analysis of LIMK2 overexpressed cells. GAPDH served as a control. K Indicated cells were subjected to soft agar assays and the relative colony numbers were quantified (n = 3, independent experiments). L KYSE450 with or without LIMK2 knock down cells were injected into nude mice (n = 10). M, N Tumor growth curves and tumor weight were evaluated between shLIMK2 and Mock group. O Representative images of esophageal at necropsy. P, Q Tumor numbers and weight to length ratio were showed (WT + Vehicle, n = 6; WT + 4NQO, n = 9; LIMK2-CKO + Vehicle, n = 6; LIMK2-CKO + 4NQO, n = 9). All data were shown as means ± SD and were analyzed with the two-tailed Student’s t test.*p < 0.05, **p < 0.01, ***p < 0.001.