Fig. 2: IL-17A signaling upregulates expression of the histone demethylase, JMJD3, via a TRAF6/NFκB pathway.
a Jmjd3 qPCR analysis of unstimulated (white) and IL-17A-stimulated (blue) keratinocytes. n = 3 biological replicates, p = 0.0021. n = 3 independent experiments. b Western blot of JMJD3 expression in unstimulated (white) and IL-17A-stimulated (blue) keratinocytes. n = 3 technical replicates, p = 0.0224. n = 3 independent experiments. Representative densitometry plot shown. c Jmjd3 qPCR analysis of Il17rafl/fl K14cre+ (red) and Il17rafl/fl K14cre- (yellow) keratinocytes. n = 3 biological replicates, p = 0.0008 (cre- medium vs. stimulation), p = 0.9131 (cre+ medium vs. stimulation), p = 0.0005 (cre- stimulation vs. cre+ stimulation), n = 3 independent experiments. d Jmjd3 qPCR analysis of Traf6fl/fl K14cre+ (red) and Traf6fl/fl K14cre- (yellow) keratinocytes. n = 3 biological replicates, p = 0.0036 (cre- medium vs. stimulation), p = 0.9007 (cre+ medium vs. stimulation), p = 0.0095 (cre- stimulation vs. cre+ stimulation), n = 3 independent experiments. e Jmjd3 qPCR analysis of keratinocytes treated with DMSO only (white), with IL-17A alone (blue), or with IL-17A and IKK inhibitor VII (2 µM) (red). n = 3 biological replicates, p = 0.0156 (DMSO vs. IL-17A), p = 0.0198 (IL-17A vs. IL-17A and inhibitor), n = 3 independent experiments. f Schematic of proposed IL-17A/TRAF6/NFκB-mediated regulation of Jmjd3. Data were analyzed for variances, and 2-tailed Student’s t tests for (a, b) and 1-way ANOVA tests for (c–e) were performed. Data are presented as the mean ± SEM.
