Fig 1: Isoliquiritin apioside does not affect the adhesion ability and expression of EMT-related proteins and integrins of HT1080 cells. (A) The wells of 96-well culture plates were coated overnight at room temperature with 5 μg/mL fibronectin (Sigma), 0.3% type I collagen solution (Cellmatrix type I-A; Nittazerachin Co., Osaka, Japan), and 10 μg/mL vitronectin (Sigma) in a volume of 50 μL. After blocking with 200 μL RPMI containing 3% BSA, ISLA-treated or -untreated HT1080 cells suspended in serum-free RPMI (1 × 105/200 μL) were added to ECM-coated wells and then allowed to adhere for 1 h at 37°C. Unbound cells were washed with PBS three times, and the attached cells were fixed and stained with 0.2% crystal violet/20% methanol (w/v) solution. After washing with distilled water, dye was lysed with 1% SDS and measured absorbance at 560 nm using SpectraMaxi3 Multi-mode reader. Data are expressed as means ± SD (n = 3). (B,C) Cells were treated with or without ISLA for 24 h and the whole cell lysates were extracted. The levels of EMT-related proteins and integrins were measured by western blotting using EMT Antibody Sampler Kit (Cell Signaling Technology, cat. no. 9782) and Integrin Antibody Sampler Kit (Cell Signaling Technology, cat. no. 4749).