Fig. 1. Effects of the RGD-to-RGE in fibulin-5 in 4T1 cells. A. Sequencing electropherograms showing the substitution of C to G in the cDNA (indicated with an asterisk), resulting in a change D56E in the amino acid sequence of fibulin-5. B. Western-blot showing the expression of fibulin-5 in 4T1 cells. Control cells transfected with an empty vector are indicated as C, interfered cells as Fibulin-5KD, transfected with the full-cDNA for fibulin-5 as Fibulin-5WT, and cells transfected with cDNA for fibulin-5 containing the RGD-to-RGE are indicated as Fibulin-5 RGE. Molecular weight is indicated on the right. β-actin was employed as loading control. Intervening irrelevant lanes are not shown. C. Morphological changes in fibulin-5 (Fibulin-5KD) interfered 4T1 cells and in 4T1 cells containing the RGD-to-RGE change in fibulin-5. Bar, 100 µm. D. Western-blot showing the expression of the EMT markers E-cadherin, N-cadherin and vimentin in the indicated cells. Molecular weight markers are indicated on the right and β-actin was employed as loading control.