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Blood, Vol. 112, Issue 3, 619-625, August 1, 2008
Integrin  E(CD103)β7 influences cellular shape and motility in a ligand-dependent fashion
Blood Schlickum et al.
112: 619
Supplemental materials for: Schlickum et al
Files in this Data Supplement:
- Figure S1. DETC (JPG, 78.8 KB)
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(A) Dendritic epidermal T cells (DETC) were isolated from epidermal sheets of wildtype ( E(CD103)+/+) or E(CD103)-deficient mice by enzymatic digestion, isolated by FACS using a CD3 -specific mAb (inserts in each panel demonstrate fluorescence of the sorted cells after isolation), and allowed overnight to re-express surface antigens and adhere to E-cadherin-coated culture dishes in the presence of a control antibody (control) or function-blocking antibodies directed against E(CD103),  7 or E-cadherin. Non-adherent cells were washed off by gentle rinsing in PBS. The formation of cell protrusions was visualized by phase contrast microscopy. Arrows indicate examples of cell protrusions/filopodia. Scale bar = 20 µm. (B) Quantitative evaluation of cells with protrusions/filopodia in cultures of DETC isolated from wildtype (left group of 4 bars) and E(CD103)-deficient mice (right group of 4 bars). The cells were cultured overnight in E-cadherin-coated dishes in the presence of a control antibody or function-blocking antibodies directed against E(CD103), 7 or E-cadherin as indicated. Values represent the mean numbers of cells with protrusions/filopodia per 100 cells from six countings of 100 cells for each condition (±SD). P values were determined by two-sided t-test comparing the antibody-treated conditions with the respective controls.
- Video 1. Mock-transfected K562 moving on E-cadherin (MOV, 1.23 MB)
- Video 2. Integrin αE-open/β7-transfected K562 cells with enhanced motility on E-cadherin. 20× lens (MOV, 1.26 MB)
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This and video 1 correspond to Fig. 1.
- Video 3. Transfectant cells seeded on an epidermal sheet: control YFP transfectant, confocal fluorescence mode (MOV, 876 KB)
- Video 4. Transfectant cells seeded on an epidermal sheet: control YFP transfectant, phase contrast mode (MOV, 1.58 MB)
- Video 5. Transfectant cells seeded on an epidermal sheet: a cell transfected with active αE-open/β7, confocal fluorescence mode (MOV, 430 KB)
- Video 6. Transfectant cells seeded on an epidermal sheet: a cell transfected with active αE-open/β7, phase contrast mode (MOV, 1.49 MB)
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Videos 3–6 correspond to Fig. 3A.
- Video 7. Confocal stacks of K562 cells transfected with αE-open/β7 and seeded on cryostat-cut sections of murine skin to visualize dendrites extended towards epidermal cells (MOV, 524 KB)
- Video 8. Corresponding phase-contrast stacks of K562 cells transfected with αE–open/β7 and seeded on cryostat-cut sections of murine skin to visualize dendrites extended towards epidermal cells (MOV, 872 KB)
- Video 9. Confocal stacks of K562 cells transfected with αE–open/β7 and seeded on cryostat-cut sections of murine skin to visualize dendrites extended towards epidermal cells (MOV, 724 KB)
- Video 10. Corresponding phase-contrast stacks of K562 cells transfected with αE–open/β7 and seeded on cryostat-cut sections of murine skin to visualize dendrites extended towards epidermal cells (MOV, 972 KB)
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Videos 7–10 correspond to Fig. 3B.
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