Oxidative Medicine and Cellular Longevity / 2013 / Article / Fig 6

Research Article

Tanshinol Attenuates the Deleterious Effects of Oxidative Stress on Osteoblastic Differentiation via Wnt/FoxO3a Signaling

Figure 6

Tanshinol attenuates the activation of FoxO3a in response to oxidative stress. (a) C2C12 cells were treated as described in Figure 4, and protein expressions from whole cell lysates and nuclear fractions were immunoblotted for FoxO3a, β-actin (whole lysates marker), and Histone H3 (nuclear marker). Representative immunoblots were shown in upper panel. Quantitative analysis of relative band intensities of protein was showed in lower panel. (b and c) Cells were transfected with the FoxO3a-luc reporter plasmid, and the treatment of Wnt3a was replaced with LY294002 (50 μM), LiCl (10 mM), or Dkk1 (500 ng/mL), and other procedures of experiments were addressed as described in Figure 5(b). (d) C2C12 cells were treated as described in Figure 4, and the expression levels of Gadd45 and CAT mRNA were quantified by qRT-PCR and normalized to GADPH mRNA. Note: (1) Con (vehicle control); (2) H (H2O2); (3) T (Tanshinol); (4) H + T (H2O2 + Tanshinol); (5) H + R (H2O2 + Resveratrol). Error bars indicate mean ± SEM of at least three independent experiments. versus vehicle control and versus H2O2 treatment.

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