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. 2006 Aug 21;26(20):7791–7805. doi: 10.1128/MCB.00022-06

FIG. 6.

FIG. 6.

Influence of cholesterol-enriched plasma membrane regions on BMP signaling pathways. C2C12 cells were treated with 50 μM lovastatin and 50 μM mevalonate and stimulated with BMP-2 as described in the text. (A) Western blot (WB) analyses using an anti-phospho- Smad1/5 (P-Smad1/5) antibody show that Smad1/5 phosphorylation is not altered by LM. For a loading control, the membrane was incubated with anti-β-actin antibody. (B) C2C12 cells were transfected with p(BRE)4-luc and pRLTK before metabolic inhibition using LM was accomplished. BMP signaling is only slightly reduced by LM. Error bars represent standard deviations from three different assays. (C) RT-PCR of ALP expression levels shown as the level of induction (n-fold) upon BMP-2 stimulation; cholesterol depletion by LM results in a reduction of the ALP transcript by 80%. Error bars represent standard deviations (SD) from duplicates. The experiment was done two times. (D) ALP activity measurements after cholesterol depletion by LM (this experiment was performed in parallel with the experiment whose results are shown in panel C) show reduction by 75%. Error bars represent SD from two independent experiments, each with triplicates. w/o, without.

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