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. 2020 Oct 9;18(6):1437–1449. doi: 10.1038/s41423-020-00559-7

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — BLT1^hi DCs migrate toward LTB₄ and accelerate dermatitis. a, b A chemotaxis assay was used to assess the migration of BLT1^hi DCs (left) and BLT1^lo DCs (right) derived from BMDCs toward LTB₄. Cells were placed in the bottom chamber, and 100 nM LTB₄ was added to the head chamber. Representative images are shown (a, upper panels). Tracking charts for each cell type are shown (n > 20). Black tracking lines indicate the cells moving forward, and red tracking lines indicate the cells moving backward (a, bottom panels). Velocity and straightness are shown in b (n > 20; error bars indicate the S.E.M.). c The fatty acid-enriched fraction from mouse ears was analyzed for LTB₄ by LC-MS/MS. The time after elicitation is shown. (−): no elicitation. d Western blot analysis for the LTA₄H protein was performed with BMDC protein lysates (WT wild-type, HE heterozygous, KO homozygous knockout). e The A23187 (calcium ionophore)-dependent production of LTB₄ by BMDCs was analyzed by LC-MS/MS. WT wild-type, HE hetero, KO knockout. f H&E-stained ear sections from LTA₄H WT and LTA₄H KO mice are shown. Bars, 100 μm. Dotted lines indicate ear thickness. g Ear thickness was measured at 0, 24, and 48 h after elicitation in WT and LTA₄H KO mice (n = 7). h The schematic shows the mouse ltb4r1 gene locus targeted to generate DC-conditional BLT1 knockout mice. i Genotyping was performed to evaluate the deletion of the targeted loci. The primer position is indicated in h. j The expression of BLT1 in splenic DCs was evaluated. k, l, m BLT1^fl/fl and BLT1 DC cKO mice were sensitized and challenged with the hapten DNFB. k H&E-stained ear sections from hapten-induced BLT1^fl/fl (left panels) and BLT1 DC cKO mice (right panels) are shown. Forty-eight hours after challenge, ear tissue was collected and stained. Bars, 100 μm. l Ear thickness was measured at 0, 24, and 48 h postelicitation in WT and BLT1 DC cKO mice (n = 6). m The mRNA expression of IFN-γ in ear tissue at 48 h postelicitation was measured by QPCR. β-actin was used as an internal control. *P < 0.05; ***P < 0.001; unpaired Student’s t-test (b, j, m); two-way ANOVA with Bonferroni’s post hoc tests (c, g, l)