Fig. 3: RANKL downregulated CCR5 expression in preosteoclast cells. | Cell Death & Disease

Fig. 3: RANKL downregulated CCR5 expression in preosteoclast cells.

From: CCL4 enhances preosteoclast migration and its receptor CCR5 downregulation by RANKL promotes osteoclastogenesis

Fig. 3

a Preosteoclast cells were cultured for 4 days in the presence of recombinant M-CSF (30 ng/ml) with RANKL (50 ng/ml) and CCL4 (10 ng/ml). Total RNA was extracted and converted to cDNA, which was used for conventional RT-PCR and (b) real-time RT-PCR. c FACS analysis. Preosteoclast cells were pretreated with (a) M-CSF (30 ng/ml), (b) M-CSF (30 ng/ml) + CCL4 (10 ng/ml), or (c) M-CSF (30 ng/ml) + RANKL (50 ng/ml) for 48 h. d Western blot of CCR5 with treatment of RANKL or CCRL4-treated cell. e Immunofluorescence staining for nuclear (blue) and CCR5 (red) in macrophage cells. f Cells were stimulated with RANKL alone (50 ng/ml) or RANKL with CCL4 (10 ng/ml) for 0–24 h. The expression levels of CCR5 mRNA were measured by real-time RT-PCR and normalized to GAPDH. The data are presented as the mean ± S.D. (n = 3; *P-value < 0.05, two-way ANOVA)

Back to article page