Figure 4 | Scientific Reports

Figure 4

From: Lycorine suppresses RANKL-induced osteoclastogenesis in vitro and prevents ovariectomy-induced osteoporosis and titanium particle-induced osteolysis in vivo

Figure 4

LY inhibited osteoclastogenesis by impairing RANKL-induced MAPK pathways without affecting the NF-κB pathway in vitro.

(A,F) BMMs were treated with or without 0.4 μM LY for 4 h and then treated with 100 ng/mL RANKL (RL) for the indicated periods. Cell lysates were analysed using western blotting. The expression of phosphorylated ERK, P38, JNK, total ERK, P38 and JNK, IκB-α (A), phosphorylated MEK1/2, MKK6, MKK7 and total MEK1/2, MKK6, MKK7 and β-actin was evaluated; (B–E) The grey levels of phosphorylated ERK, P38 and JNK were quantified and normalized to total ERK, P38 and JNK using Image J. The grey level of IκB-α was normalized to β-actin (*P < 0.05; **P < 0.01). (G–I) The grey levels of phosphorylated MEK1/2, MKK6 and MKK7 were quantified and normalized to total MEK1/2, MKK6 and MKK7 using Image J (*P < 0.05; **P < 0.01). (J) BMMs were treated with or without 0.4 μM LY for 4 h and then treated with or without 100 ng/mL RANKL for 5 min. Cell lysates were analysed using western blotting. The expression of phosphorylated TAK1, total TAK1 and β-actin was evaluated. (K) The grey levels of phosphorylated TAK1 were quantified and normalized to β-actin using Image J. (L) BMMs were treated with RANKL, with or without 0.4 μM LY, for the indicated periods. Cell lysates were analysed using western blotting. The expression of NFATc1 and β-actin was evaluated. (M) The grey levels of NFATc1 were quantified and normalized to β-actin using Image J (*P < 0.05). (N) Schematic diagram of the mechanism by which LY inhibits osteoclast differentiation and function.

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