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. 2019 Jan 4;9:3050. doi: 10.3389/fimmu.2018.03050

Figure 3.

Figure 3

Inhibition of the IRE1α endonuclease domain by the aldehyde 4μ8C does not affect BMDC cellularity or expression of costimulatory molecules, but reduces the production of cytokines upon MEL stimulation. (A) FL-DCs were stimulated with increasing doses of 4μ8C or DMSO and composition of DC subtypes was monitored 24 h post treatment. (B,C) FL-DCs were pretreated with 20 μM 4μ8C or DMSO for 2 h and stimulated with 100 μg/ml MEL for additional 16 h. Expression of MHC-II, CD80, CD86, and PD-L1 were measured by flow cytometry. Histograms shown in (B) are one representative experiment out of five of cDC1 FL-DC (CD24+) and cDC2 FL-DC (SIPRα+) generated in cultures and graphed in (C). (D) FL-DCs were pretreated with 20 μM 4μ8C or DMSO for 5 h and stimulated with 100 μg/ml MEL for additional 16 h. TNF-α, IL-6, and IL-10 were quantified by cytometric bead array. (E) FL-DCs were pretreated with 50 μM 4μ8C or DMSO for 5 h and stimulated with 100 μg/ml MEL for additional 16 h. IL-12p40 was analyzed by intracellular staining. Contour plots and graphs are for cDC1 FL-DC (CD24+) generated in cultures. For (D,E), each symbol in the graphs represents data derived from one independent experiment. For all error bars represent mean ± SEM. *p < 0.05, ***p < 0.001 (paired Student's t-test).