FIGURE 7.

Extracellular mutant cochlin binds to primary fibrocytes and reduces cell survival. A, fibrocytes (FC) from WT mice were isolated and cultured in vitro and treated with CM (1:5 dilution) collected from transiently transfected HEK293T cells expressing vehicle control (Vec) or WT or mutant G90E cochlin. After treatment, the supernatant was removed, and cells were washed twice with phosphate-buffered saline. The cell lysates were analyzed by 15% reducing SDS-PAGE, followed by Western blotting using anti-LCCL antibody. B, primary fibrocytes were incubated with CM from 293T cells expressing vehicle (Con), WT cochlin alone, WT and mutant G90E cochlins together (W/M), or mutant G90E cochlin alone (Mut) for 72 h. Cell survival was measured using the MTS assay. *, p < 0.05. FL-Coch, full-length cochlin.