Figure 1

Knock-down of ITGA6 affect adhesion of human thymic epithelial cells. TEC were transfected either with siRNA oligonucleotides targeting the ITGA6 gene (ITGA6 siRNA) or with a scramble siRNA oligonucleotide sequence (Control siRNA). Results of TEC treated either by using control siRNA or ITGA6 siRNA are presented in each assay. ITGA6 gene expression was measured in five independent transfection experiments by qRT-PCR, seen in panel a. The protein encoded by ITGA6 gene, CD49f, was evaluated on the surface of TEC by cytofluorometry after gene silencing experiments (panel b). Gray histogram represents control isotype, the filled histogram depicts the cells transfected with ITGA6 siRNA and the hachured are the cells with control siRNA. CD49f expression was also determined by immunofluorescence, as shown in panel c. In all cases, gene expression and the corresponding integrin α-chain were consistently and significantly impaired. Transfected TEC were plated onto laminin coat (d). Note that the control cells (control siRNA) attached and spread well than knocked cells (ITGA6 siRNA). The cell number in both condition were quantified (e). The individual bars represent mean ± standard error (n = 5). Magnification bars correspond to 35 µm in panel c and 70 µm in panel d. All experiments were done 48 hours of gene silencing and using a siRNA dose of 5nM. **p < 0.01.