Figure 5
Bzb does not alter expression of Wnt ligands, receptors, and Wnt antagonists or OPG protein in OBs. Saos-2 cells were cultured in the medium in the absence or presence of 10 nM of Bzb for 24 hours. Total RNA was isolated, and RT-PCR analysis was performed to measure mRNA expression of WNTs (A), FZDs (B), and LRP5/6 (C), as described in “RT-PCR analysis.” The Saos-2 (D,E) cells were transfected with TOPflash plasmid DNA. The cells were treated with indicated concentrations of Bzb in the presence or absence of Wnt3a (100 ng/mL, D) or lithium chloride (10 mM, E) for 24 hours. Lysates was harvested and subjected to luciferase quantification. The C2C12 (F) or C3H10T1/2 (G) cells were treated in the presence or absence of Bzb (100 nM) or Wnt3a (100 ng/mL) for 48 hours and supernatants harvested for ELISA analysis of OPG protein concentrations. Results are mean plus or minus SD (n = 3). **P < .01, ***P < .001 vs controls.

Bzb does not alter expression of Wnt ligands, receptors, and Wnt antagonists or OPG protein in OBs. Saos-2 cells were cultured in the medium in the absence or presence of 10 nM of Bzb for 24 hours. Total RNA was isolated, and RT-PCR analysis was performed to measure mRNA expression of WNTs (A), FZDs (B), and LRP5/6 (C), as described in “RT-PCR analysis.” The Saos-2 (D,E) cells were transfected with TOPflash plasmid DNA. The cells were treated with indicated concentrations of Bzb in the presence or absence of Wnt3a (100 ng/mL, D) or lithium chloride (10 mM, E) for 24 hours. Lysates was harvested and subjected to luciferase quantification. The C2C12 (F) or C3H10T1/2 (G) cells were treated in the presence or absence of Bzb (100 nM) or Wnt3a (100 ng/mL) for 48 hours and supernatants harvested for ELISA analysis of OPG protein concentrations. Results are mean plus or minus SD (n = 3). **P < .01, ***P < .001 vs controls.

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