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. 2019 Mar 22;20(6):1467. doi: 10.3390/ijms20061467

Figure 4.

Figure 4

Zoledronate enhanced the RANKL expression via interleukin-6 (IL-6)/Janus activated kinase (JAK2)/ signal transducer and activator of transcription 3 (STAT3) pathway in MLO-Y4 cells. (a,b) MLO-Y4 cells were cultured in the presence of indicated doses of clodronate (C; 0.1 or 1 µM) or zoledronate (Z; 0.1 or 1 µM) for 2 days and the mRNA expressions of IL-6 and gp130 were examined by quantitative real-time PCR; (c) MLO-Y4 cells were cultured in the medium containing zoledronate (1 µM) for 2 days and serum-starved for 5 h. The cells were pretreated with vehicle (DMSO) or AG490 (20 µM) for another 1 h and re-stimulated with zoledronate. The phosphorylation of STAT3 was evaluated by immunoblotting (upper). The relative band intensity of p-STAT3 was normalized by STAT3 with densitometry and the quantitative results are shown as graphs (lower); (d,e) MLO-Y4 cells were cultured in the presence of zoledronate (1 µM) together with AG490 (0, 10, and 20 µM) for 2 days. After the culture, the expression of RANKL was determined by quantitative real-time PCR (d) or immunoblotting (e). Quantitative data are presented as mean ± SD (* p < 0.05).