Journal: Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology

Article Title: Rho and Rac, but not ROCK, are required for secretion of human and mouse eosinophil-associated RNases

doi: 10.1111/cea.13292

Figure Lengend Snippet: CCL11-mediated EAR secretion from eosinophils requires Rho activity. RNase activity was measured in supernatants of mCCL11-stimulated mouse (A) and hCCL11-stimulated human (B) eosinophils in the presence of Rho GTPase blocker, C3-transferase, as described in Material and Methods. Data are mean ± SD of five independent experiments (A) or mean ± SD from triplicate wells and are representative of four independent experiments (B). Asterisks represent P values < 0.05 (*), or < 0.01 (**).

Article Snippet: In some experiments, cells were pretreated for two hrs at 37°C with the Rho blocker, C3 transferase (5 and 10 μg/ml, Cytoskeleton Inc., Denver, CO, USA), or for 20 min at 37°C with the vesicle formation inhibitor, brefeldin A (10 μM, Enzo Life Sciences LTD., Exeter, UK), the ROCK inhibitor, Y27632 (30, 50 and 100 μM, Calbiochem-EMD), the Rac inhibitor, EHT 1684 (5 and 10 μM, Tocris Bioscience, Bristol, UK) and the microtubule polymerization blocker, nocodazole (10 and 100 μM, Calbiochem-EMD), before and during chemokine stimulation.

Techniques: Activity Assay