Journal: Journal of Leukocyte Biology

Article Title: An essential role for Rab27a GTPase in eosinophil exocytosis

doi: 10.1189/jlb.0812431

Figure Lengend Snippet: (A) RT-PCR analysis of differentiated HL-60c15 cells and bmEos cultured from C57BL/6 mice. (B) Western blot analysis of Rab27a expression in eosinophils from mouse and human sources. BmEos were isolated from C57BL/6 mice (left) and bmEos, and peripheral blood (PB) eosinophils were prepared from IL-5 Tg mice (right), to compare Rab27a expression with that in human eosinophils and differentiated HL-60c15 cells. A total of 20 μg protein was loaded in each lane. (C) Subcellular fractionation of peripheral blood eosinophils from IL-5 Tg mice. An enzymatic assay was used to detect EPX in fractions containing crystalloid granules (fractions 6–10), and LDH was assayed to identify cytosolic fractions (fractions 16–23). Western blot analysis showed Rab27a (fractions 6–13) and gp91phox (a marker for plasma membrane, fractions 11–15).

Article Snippet: Preparation of eosinophils HL-60 clone 15 (HL-60c15) cells (ATCC, Manassas VA, USA), chosen for this study based on their propensity to differentiate into highly granulated eosinophil-like cells, were treated with 500 μM n -butyrate [ 26 ].

Techniques: Reverse Transcription Polymerase Chain Reaction, Cell Culture, Western Blot, Expressing, Isolation, Fractionation, Enzymatic Assay, Marker