ZOL specifically attenuates RANKL-induced NF-κB and JNK signalling. (A) RAW264.7 cells were induced with RANKL for the indicated times following pre-treatment with ZOL (5 µM) for 4 h. Western blot analysis was performed using specific antibodies. The band intensities were quantified using Image J software. The ratios of band intensity of (B) p-IκBα/IκBα, (C) p-p65/p65, (D) p-JNK/JNK, (E) p-p38/p38 and (F) p-ERK/ERK are shown. (G) ZOL inhibited osteoclast differentiation at an early stage. RAW264.7 cells were treated with or without 5 µM ZOL at days 0, 1, 3 and 4. They were subsequently treated with 100 ng/ml RANKL for 5 days, and then subjected to TRAP staining. (H) The number and (I) area of osteoclasts were quantified. *P<0.05, **P<0.01 vs. the RANKL-only group. TRAP, tartrate-resistant acid phosphatase; ZOL, zoledronic acid; RANKL, receptor activator of nuclear-κB ligand.