Fig. 4

D10 cells treated with 4μ8c produce IL-5, but they cannot secrete it. D10 cells were treated as in Fig. 3a. a At 20 h, the cells were treated with monensin for an additional 4 hours and intracellular staining was performed for IL-5 and IL-13. b The supernatants were isolated at 20 h from the cells shown in A and an ELISA was performed in duplicate for IL-4, IL-5, and IL-13. The data is representative of six experiments. c The average percent of cells positive for IL-5 and IL-13 are graphed from six experiments. The error bars represent the standard error of the mean. Hypothesis testing was done by Student’s T test unpaired, Welch’s correction (p value < 0.05). d A cytokine secretion assay was performed on D10 cells treated as above. The data shown is a representative of the percent IL-5 secreting cells from each sample group. The experiment was done with duplicate samples three times. e The percent IL-5 secreting cells for each treatment that was performed in duplicate was averaged and graphed for the three individual experiments performed. f Protein was isolated from cells treated as in 3A and immunoblotted with IL-5, IL-13, and β-actin antibody. The data is representative of two experiments