The pattern of chemokine and ICAM-1 expression in LECs depends on the nature of the inflammatory stimulus. (A) Quantitative PCR analysis performed on amplified cDNA from ex vivo isolated LECs revealed marked differences in the pattern of ICAM-1 and chemokine production in response to CHS- or CFA-induced skin inflammation. Pooled data from 3 or 4 independent experiments are shown. (B) Up-regulation of ICAM-1 in LECs (gated on CD45⁻CD31⁺podoplanin⁺ cells) in response to CHS- but not CFA-induced inflammation was confirmed by FACS analysis performed on ear tissue single-cell suspensions. ΔMFI indicates difference in the median fluorescent intensity between the ICAM-1 signal and the isotype signal. Black line represents ICAM-1 staining; and gray, tinted line, isotype control staining. (C-E) Conditionally immortalized Immorto LECs (imLECs) were treated with combinations of IFN-γ/TNF-α or MDP/LTA to mimic CHS- or CFA-induced inflammation, respectively. (C) FACS analysis of the induction of ICAM-1 and VCAM-1 by the treatment. (D) Analysis of the mRNA chemokine expression profiles induced by the treatment. (E) Chemokines were measured in the cell culture supernatant of imLECs treated with IFN-γ/TNF-α or LTA/MDP using a FACS-based cytokine assay. *P < .05. **P < .01. ***P < .001.