Extended Data Fig. 3: Depletion of spermine impedes granule assembly.
From: Mast cell extracellular granules are bioactive condensates assembled by heparin and polyamine
a, Spermine levels in APCHA-treated cells were determined by flow cytometry. Dashed line depicts the spermine-positive gate. b, Validation of avidin bead-based MCEG quantification method. Serially diluted MCEGs were incubated with equal numbers of avidin beads for 1 h at room temperature. The number of MCEGs prior to avidin bead incubation was determined by flow cytometry. The number of MCEGs per bead was plotted against the dilution factor. c, Western blot analysis of mast cell mediators in whole cell lysate from inhibitors-treated BMMCs. Band intensities for each mediator were quantified and normalized to β-actin. All normalized values were expressed as fold change relative to the vehicle control and plotted in the right panels (each data point represents one independent experiment, n = 3). P value by ordinary one-way ANOVA. d, Beta-hexosaminidase levels in different cell fractions as separated by centrifugation (upper panel) were expressed as percentages of total β−hexosaminidase in the cells. N = 4 biological replicates. Data are presented as Mean ± SD. One representative of three independent experiments was shown.
