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

Fig 4. TRAFD1 knockdown inhibits resorbing activity of osteoclasts.

Fig 4

(A) Stable knockdown of TRAFD1 was achieved by transduction with lentivirus expressing shRNA (or scrambled control) in RAW264.7 cells. Clones were selected with different degrees of knockdown in the absence of RANKL, as shown (clones 1.7, 2.3; and 2.6). Q-PCR experiments were performed at least 3 times and representative graph shows means+ s.d. of duplicates. *** P<0.0001 versus control group. (B) Resorption assay was performed with the knockdown clones in A. Cells were cultured on 24-well Osteo Assay (HA) plates for 10 days in the presence of RANKL (10 ng/ml). Cells were removed and the plate was scanned at high resolution. Representative wells are shown on lower panel (black = resorbed area). The percentage of resorbed area over total area of well is indicated on upper panel. Analysis of resorbed area was measured by Image J software. One-way ANOVA was carried out and values are mean +s.d. of n = 3 independent experiments analyzing 3 wells/condition. *** P<0.0001 versus control group.