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. 2015 May 19;10(5):e0127537. doi: 10.1371/journal.pone.0127537

Fig 8. TRAFD1 is required for TRAP and cathepsin K secretion.

Fig 8

(A) Differentiated RAW 264.7 cells and shTRAFD1 cells were grown for 5 days on Osteo Assay plates in the presence of RANKL. Conditioned media were collected 2 and 5 days post differentiation and TRAP secretion was measured biochemically. A representative graph is shown. One-way ANOVA was carried out and values are mean +s.d., n = 8 per time point. *** P<0.0001 vs. controls. (B) TRAFD1 knockdown and control RAW264.7 cells were cultured on Osteo Assay plates in the presence of RANKL. Culture supernatant (sup) was collected 24 hours post differentiation. 40 μl of conditioned media were subjected to western blot analysis for the presence of cathepsin K. Total cell lysates (TCL) from each well served as total protein controls. The same gel was re-probed with lamin B1 antibody. The experiments were performed 3 times and representative gels are shown.