Journal: Oncotarget

Article Title: miR-135a inhibits tumor metastasis and angiogenesis by targeting FAK pathway.

doi: 10.18632/oncotarget.16098

Figure Lengend Snippet: Figure 2: FAK is a target gene of miR-135a. (A) The binding site of miR-135a in the FAK 3′UTR and the skeleton of FAK 3′UTR luciferase reporter vector. (B) The protein expression of FAK, p-ERK, ERK, ROCK1, p-LIMK1 and LIMK1 in SGC-7901 and BGC-823 stable cell lines are examined by Western Blot. (C) Relative expression of secreted VEGFA in the culture supernatant of cancer cells is detected by ELISA assay. (D) The protein expression of phosphorylated FAK (pTyr397) in cells that treated with conditioned medium from control or miR-135a overexpressing stable cell lines. (E) miR-135a overexpressing cells were transfected with 200 nM miRNA inhibitor or control RNA, then FAK protein level was examined with Western blot assay. (F) The effect of miR-135a mimics on the luciferase activity of FAK 3′UTR wild type (WT) and deletion type (DEL) are detected by luciferase reporter assay. * p < 0.05

Article Snippet: FAK1 (Cell signaling technology, USA), ROCK1 antibody (Origene, USA), p-LIMK1 (Cell signaling technology, USA), GAPDH antibody (Kangchen, China), p53 (Santa Cruz, USA) and Flag (or DKK) antibody (Abmart, China) were used.

Techniques: Binding Assay, Luciferase, Plasmid Preparation, Expressing, Stable Transfection, Western Blot, Enzyme-linked Immunosorbent Assay, Control, Transfection, Activity Assay, Reporter Assay