Panel A, C3H10T1/2 cells were treated either with vehicle, 15 ng/ml recombinant purified Wnt3a, 5 ng/ml recombinant purified TGF-β1, or with both ligands as indicated for 7 days as indicated. Panel B, within 4 days of treatment, 15 ng/ml Wnt3a significantly upregulates SM22α mRNA accumulation. Panel C, western blot analysis confirms that Wnt3a increases SM22α protein levels as well (see also Figs. 1E and 2B). Duplicate blots are shown, representative of the 4 independent blots analyzed by digital image analysis. Unlike Wnt3a, Wnt1 and Wnt5a do not upregulate SM22α mRNA accumulation (panel D) or SM22 protein levels (panel E). All Wnt treatments were at 15 ng/ml.