Fig 1: Ectopic expression of ANK3 exerted anti-oncogenic role in PTC. (A) Transfected with ANK3 expression vectors in B-CPAP and TPC-1 cells, ANK3 expression was significantly increased compared to the control group (empty vector cells). (B–D) Ectopic expression of ANK3 significantly reduced the cell proliferation and anchorage-dependent growth with MTT assays and colony formation assays. (E, F) Ectopic expression of ANK3 suppressed cell invasion of B-CPAP and TPC-1 cells. (G, H) Flow cytometry showed that ectopic expression of ANK3 increased cell apoptosis. (I–L) Ectopic expression of ANK3 significantly suppressed cell migration of B-CPAP and TPC-1 cells. (*P<0.05, Student’s t-test, n=3 independent experiments).
Fig 2: ANK3 expression in tumour tissues was detected in a tissue array by using immunohistochemistry. (A–D) ANK3 was dominantly expressed in cytoplasm close to the membrane of PTC cells. The above:100×; The below: 200×. (E, F) ANK3 expression in tumour tissues and non-tumour tissues was detected in a tissue array by using immunohistochemistry (100×). Lower figure showed ANK3 staining has been detected in cytoplasm of normal thyroid follicle epithelial cells and colloid of thyroid follicle. ANK3 staining was also dominantly expressed in cytoplasm close to the membrane of PTC cells (200×).
Fig 3: (A) Transfection of ANK3 in both B-CPAP and TPC-1 cells significantly increased E-cadherin expression in the membrane of PTC cells by immunofluorescence assay. (B–E) Ectopic expression of ANK3 was obviously enhanced E-cadherin protein expression and significantly attenuated vimentin, snail, and twist protein expression in both B-CPAP and TPC-1 cells. (Con: cells transfected with empty vector; OE: cells transfected with ANK3 expression vector).
Fig 4: Ectopic expression of ANK3 suppressed PTC tumorigenesis. (A, B) Representative image of tumors in xenograft mouse model. (C, D) Over-expression of ANK3 TPC-1 cells dramatically inhibited tumor growth in the nude mice. Tumor volumes and weight were measured and presented as mean ± SD (n=5 mice per group). P values were determined by the one-way analysis of variance (ANOVA). (E) TPC-1 xenografts overexpressing ANK3 decreased cell proliferation by Ki-67 staining and augmented E-cadherin expression. (*P<0.05).
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