Altered adhesion molecule expression and reduced leukocyte adhesion in TNFR1^−/− mice. (A) mRNA expression of TNFR1 in mouse heart ECs, brain ECs, and mouse ear (data were normalized to aldolase expression; relative mRNA expression was calculated by dividing TNFR1 mRNA expression of TNFR1^+/+ mice, mouse heart ECs, and brain ECs though the background TNFR1 mRNA in TNFR1^−/− mice, corresponding to the detection limit, set as 1). (B) mRNA expression of P-selectin, E-selectin, ICAM-1, and VCAM-1 in TNFR1^−/− mice and TNFR1^+/+ mice, 4 hours after elicitation of DTHR (data were normalized to aldolase expression). Data are given in a decade log scale (n = 9 or 10). (C) Immunofluorescence staining of P-selectin, E-selectin, ICAM-1, and VCAM-1 (red) 2.5 hours and 4.0 hours after TNCB challenge in TNFR1^+/+ and TNFR1^−/− mice (green represents nuclei; blue represents type IV collagen; n = 3). (D) Leukocyte adhesion to vascular endothelia in TNFR1^+/+ and TNFR1^−/− mice 3.5 hours after elicitation of DTHR as determined by intravital fluorescent microscopy (n = 6 or 7). (E) Noninvasive intravital microscopy images of leukocyte adhesion to vascular endothelia in TNFR1^+/+ and TNFR1^−/− mice, 4.5 hours after elicitation of DTHR (supplemental Videos 1,2). (F) Leukocyte rolling and (G) firm adhesion of rhodamine-stained leukocytes to vascular endothelia in TNFR1^+/+ mice after application of P-selectin, E-selectin, ICAM-1, and VCAM-1 blocking Abs or istotype control 2.5 and 3.5 hours after elicitation of DTHR. Analysis was performed by intravital fluorescent microscopy (n = 5-7).