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. Author manuscript; available in PMC: 2013 Mar 11.
Published in final edited form as: Bone. 2007 Sep 26;42(1):180–192. doi: 10.1016/j.bone.2007.09.046

Figure 8.

Figure 8

nha-oc/NHA2oc silencing inhibits osteoclast differentiation and activity.

(A) RANKL-stimulated RAW 264.7 cells were transfected with nha-oc/NHA2-specific (S1/S2 (+)) or control (Neg (+)) siRNA. We first tested two siRNA concentrations: 50 nM (lanes 50) or 100 nM (lane 100) and chose the former for further experiments. Total RNA was isolated and subjected to RT-PCR analysis using nha-oc/NHA2- (top) or β-actin (bottom)- specific primers. Image analysis shows that the knock-down efficiency is over 90%.

(B) (C) RANKL-stimulated RAW 264.7 cells were transfected with nha-oc/NHA2-specific (S1/S2 +) or control (Neg +) small inhibiting RNA (siRNA). After 4 days cells were fixed and stained for TRAP. The formation of multinucleated TRAP+ cells (white arrow-heads) is significantly reduced in nha-oc/NHA2-siRNA treated cultures (bottom panel) * p < 0.05

(D) nha-oc/NHA2-silencing inhibits the ability of differentiated RAW 264.7 cells to form resorption pits on a calcium phosphate matrix * p < 0.05.