Fig. 6

BB-Cl-Amidine treatment suppresses cell migration and enhances cell adhesion in MCF10DCIS.com cells. a Scratch assays were performed on serum-deprived MCF10DCIS.com cells following either no treatment (Untreated) or treatment with EGF, EGF + BB-Cl-Amidine, or with BB-Cl-Amidine alone. MCF10DCIS.com cells were pre-treated with BB-Cl-Amidine for 23 h and then EGF was added for 1 h. Cells were fixed using 4% paraformaldehyde 24 h after striking the wound (24 h) and then visualized and imaged using light microscopy to determine the extent of the wound closure. Widths of initial wounds are indicated by the lines. Cells were fixed immediately after striking the wound (0 h) to indicate the size of the initial wounds. b MCF10DCIS.com cells were treated either with DMSO or 2 μM BB-Cl-Amidine. One hour after treatment, the cells were imaged using light microscopy. Scale bar = 100 μm. c Total RNA was isolated from MCF10DCIS.com cells treated with either 0 (DMSO), 1 μM, or 2 μM BB-Cl-Amidine. CDH1 mRNA levels were determined by qRT-PCR (SYBR) using DMSO treated control cells as a reference and β-actin for normalization. Data were analyzed using the 2 ^{-ΔΔ C(t)} method and are expressed as the mean ± SD from two biological replicates with three technical replicates per biological replicate (* p < 0.05; ** p < 0.005). d Anti-E-cadherin immunofluorescence staining was performed on serum-deprived MCF10DCIS.com cells following either no treatment (Untreated) or treatment with EGF, EGF + BB-Cl-Amidine, or with BB-Cl-Amidine alone. MCF10DCIS.com cells were pre-treated with BB-Cl-Amidine for 23 h and then EGF was added for 1 h. Scale bar = 50 μm