Fig. 1

CXCL10 induces migration of BMMs and CD4⁺ T cells through CXCR3 but not TLR4. a, b BMMs and CD4⁺ T cells were isolated from wild-type (WT), Tlr4 ^–/–, and Cxcr3 ^–/– mice and then the cells were serum starved. Cell migration in response to CXCL10 (100 ng/ml) was assessed in transwell chambers for 12 hours. Representative images of migrated BMMs. (a, left panel; scale bar, 200 μm). Number of migrated BMMs (a, right panel) and CD4⁺ T cells (b). c Serum-starved BMMs (left panel) and CD4⁺ T cells (right panel) were preincubated with DMSO or 10 μM PD98059 for 1 hour, and migration was measured in the absence or presence of CXCL10 (100 ng/ml) after 12 hours. d Serum-starved BMMs (left panel) and CD4⁺ T cells (right panel) were stimulated with CXCL10 (100 ng/ml) for the indicated times. Total cell lysates were immunoblotted with the indicated antibodies. e, f WT or Cxcr3 ^–/– BMMs were infected with adenovirus carrying empty vector (Ax1w1) or CA-MEK1 (AxMEK^CA). After 24 hours, the infected cells were serum starved and stimulated with PBS (vehicle) or CXCL10 (100 ng/ml) for 5 min. Total cell lysates were immunoblotted with the indicated antibodies (e). Migration of the infected cells in response to CXCL10 (100 ng/ml) was assessed in transwell chambers for 12 hours. Representative images of migrated BMMs (f, left panel; scale bar, 200 μm) and the number of migrated BMMs (f, right panel). Results shown are representative of three independent experiments (n = 3), and values are expressed as mean ± SD. *P < 0.001 by one-way ANOVA followed by Dunnett’s test. CXCL10 C-X-C motif chemokine 10, Tlr4 toll-like receptor 4, Cxcr3 CXC chemokine receptor 3, n.s. nonsignificant, BMM bone marrow-derived macrophage