Fig. 3.

Detection of poly I:C-stimulated production of intercellular adhesion molecule-1 (ICAM-1) protein. (a) HT-29 cells, 2 × 10⁵, were plated on coverslips the day before stimulation. After stimulation with or without poly I:C, the cells were stained with anti-ICAM-1 antibody followed by fluorescein isothiocyanate-conjugated goat anti-mouse ICAM-1 monoclonal antibody. Images (×63) were taken using a LSM510 confocal laser microscope (Carl Zeiss, Heidelberg, Germany). (b) HT-29 cells were stimulated with or without 100 µg/ml of poly I:C for 24 h. After stimulation, the cell lysates were prepared and ICAM-1 was immunoprecipitated with anti-ICAM-1 antibody. The immunoprecipitated samples were loaded on 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis and subjected to Western blotting. The clear 100 kDa band was detected when HT-29 cells were stimulated with poly I:C. (c) HT-29 cells were stimulated with (+) or without (−) 100 µg/ml of poly I:C for the times indicated. The culture supernatants were collected and the amount of sICAM-1 was measured by enzyme-linked immunosorbent assay. The means of four different experiments are shown.