(A) Representative image of PDPN-positive cells in culture. (B) The PDPN-positive cells were isolated from infarcted mouse hearts 2 days after MI and culture expanded in vitro. The representative flow cytometry scatter plots of the flow cytometry analysis showed that 85% of the cell population is PDPN positive and LYVE-1 negative (n = 3/isolations). (C and D) Flow cytometry analysis of the circulating PDPN-positive cells in the peripheral blood of control (C, sham operated) and 2 days after MI (D) animals. Importantly, the BM lineage–positive cells (stained with the lineage-positive cocktail antibodies) do not express PDPN and the PDPN-positive cells did not increase in the circulation after myocardial infarction, n = 3/isolations. (E) Quantitative analysis of the PDPN-positive cells that migrated from the basal to the apical side of transwell inserts under different conditions; the migratory capacity of PDPN-positive cells toward LPS-conditioned medium was significantly neutralized by 5 μg/mL of anti-PDPN antibodies. Data are presented as mean ± SEM. ****P < 0.0001 control versus LPS-conditioned medium. ^$$$$P < 0.0001 LPS-conditioned medium versus LPS-conditioned medium plus anti-PDPN Ab 1. ^####P < 0.0001 LPS-conditioned medium versus LPS-conditioned medium plus anti-PDPN Ab 2. One-way ANOVA and Bonferroni’s post hoc test were performed among all groups. Data are represented as a mean value of 3 independent experiments. For each experiment, between 3 and 6 samples per group were used.