Figure 6.

Chemokine Expression Triggered by P-407 Treatment

(A–D) The effect of P-407 on mRNA expression of chemokine/cytokine was analyzed by RT-PCR. Relative expression of (A) CCL3, (B) CCL4, (C) CCL2, and (D) IL-6 to the housekeeping gene HPRT. Kidney, liver, and heart specimens from mice treated with P-407 were compared with those from PBS-treated animals at each time point.

(E) CCL4 and CCL2 mRNA expression relative to HPRT in CD45⁺F4/80⁺ sorted tissue macrophages from liver, heart, and kidney after 14 days of P-407 or PBS treatment. Data are pooled from three animals per condition and representative of two independent experiments.

(F) Plasma concentrations of CCL4 and CCL2 after 7, 14, and 28 days of P-407 or PBS treatment. Values represent mean ± SE, n = 4 mice per group.

(G) Frequencies of Gr1^high and Gr1^low monocyte subsets in P-407-treated B6 mice injected with PT or PBS. PBS-treated mice were used as controls. Values represent mean ± SE, n = 3 mice per group. Results are representative of three independent experiments. Significant p values are indicated (^∗p < 0.05, ^∗∗p < 0.01, and ^∗∗∗p < 0.001; unpaired t test). ND, nondetectable; ns, nonsignificant.

(H) Transwell migration assay. Fluorescence-activated cell-sorted Gr1^high and Gr1^low monocytes were added to a transwell chamber for 2 hr in the presence of recombinant mouse CCL4 (1–1,000 ng/ml) or PBS (0). The number of migrated cells per field was quantified (five fields per sample); pool of n = 3 mice in triplicate. Data are mean ± SE. ^∗∗∗p < 0.001 (unpaired t test).

See also Figure S6.