Fig. 5.

Contributions to HA synthesis and turnover by cell type in mammary tumors. EpCAM+, F4/80+, and CD45-/CD90.2 + cells were isolated from (A) 4T1 (n = 3) and (B) HC11/R1-LM (n = 4) tumors using the Miltenyi Bead Isolation kit. Gene expression by qRT-PCR was analyzed utilizing a panel of HA-related genes: hyaluronan synthesis (Has1-3), hyaluronidases (Hyal1,2), and hyaluronan receptors (Cd44, Rhamm, Lyve1). (C) J774 macrophages were incubated with DMEM, HC-11/R1-LM conditioned media, or 4T1 conditioned media and lysates were assayed for hyaluronidase expression by ELISA or (D) HA activity with an HA Activity ELISA. Error bars represent standard error of the mean. P values * p < 0.05; ** p < 0.01; *** p < 0.001