Fig. 5.

IRAK-M is necessary for Il6, Csf3, Cxcl2 and Ccl5 expression in dendritic cells upon IL-33 induction. a Western blot shows IRAK-M levels in shIRAK-M expressing DC2.4 cells. Right panel shows heat map showing expression levels of different genes according to the affymetrix gene expression profiling analysis. b qRT-PCR analysis for expression of the indicated genes normalized to actin. The numbers over bars (95, 55 and 27) indicate the folds of induction of target genes in the presence vs absence of IL-33 in the sh IRAK-M group. c Quantification of IL-6 release measured by ELISA. d IRAK-M, IRAK-M S110E or IRAK-M P111A stably expressing DC2.4 cells were either treated or not with IL-33. Also as indicated, some cells were pretreated for 3 days with 5 μM or 10 μM of ATRA before IL-33 induction. e Relative gene expression of Il6, Csf3, Cxcl2 and Ccl5 in the different samples as in d normalized to actin. f IRAK-M was stably expressed in pLKO or TET on shPin1 expressing cells, followed by induction with IL-33, before western blot to examine protein expression of IRAK-M, PIN1 and tubulin as well as g qRT-PCR to determine the relative gene expression for Il6, Csf3, Cxcl2 and Ccl5 in the different samples. The data were analyzed by a Student’s two-tailed t test and the values are reported as mean ± standard errors of the means (SEM). *- statistical significance (P < 0.05), **- significance (P < 0.01)