Intracellular Secretory Leukoprotease Inhibitor Modulates Inositol 1,4,5-Triphosphate Generation and Exerts an Anti-Inflammatory Effect on Neutrophils of Individuals with Cystic Fibrosis and Chronic Obstructive Pulmonary Disease
Figure 4
SLPI inhibits neutrophil cytoskeletal rearrangements. (a) Immunoblot with anti-actin antibody for the distribution of G-actin (in the supernatant fraction, S) and F-actin (in the pellet fraction, P) in untreated cells, IL-8/fMLP (1.2 nM/1 μM), SLPI (480 nM), or control wortmannin (Wort; 100 nM) treated cells. The distribution ratios calculated using constants obtained of Western blot densitometry values are illustrated. (b) The effect of SLPI (480 nM) on talin-1 (190 kDa fragment) or vinculin cleavage (90 kDa fragment) after PMA (1.6 μM, positive control), or IL-8/fMLP activation for 10 min was analysed by Western blotting using anti-talin-1 and anti-vinculin monoclonal antibodies. Equal sample loading was confirmed by Coomassie blue stained gels (top panels) and the β-actin immunoblots (lower panels). Results of three separate experiments were expressed as relative densitometry units ((c) and (d)) and each bar is the mean ± S.E. * between SLPI treated and untreated cells for the respective stimuli calculated by Student’s -test.
