Antisense inhibition of decorin expression in myoblasts decreases cell responsiveness to transforming growth factor beta  and accelerates skeletal muscle differentiation.

Cecilia Riquelme, Juan Larraín, Elke Schönherr, Juan Pablo Henriquez, Hans Kresse, and Enrique Brandan

Pages 3593 and 3594: Figs. 4 and 6 were reversed although the legends are correct. The correct figures and legends are shown below.


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Fig. 4.   The expression of myogenin in antisense decorin clones is reverted by specific decorin re-expression and by the addition of exogenous decorin. A: left panel, myogenin expression was evaluated in wild type cells and antisense decorin-transfected cells (WT and A6) and in transfected cells infected with an adenovirus containing the full-length sequence for human decorin (WTi and A6i). Northern blot analysis for myogenin under growth conditions was performed. 10 µg of total RNA isolated from myoblasts was separated by electrophoresis, blotted onto nylon membranes, and hybridized with a 32P-labeled myogenin probe (Myo). The methylene blue-stained nylon membrane is shown in the lower part of the panel, and the ribosomal RNAs are indicated. The transcript size is indicated. Right panel, the extent of decorin synthesized by the adenovirus-infected cells described in A was evaluated by specific immunoprecipitation with antibodies against human decorin as described under "Experimental Procedures." B, A6 cells were incubated for 48 h with decorin purified from bovine cartilage (left panel) or from mouse skeletal muscle (right panel). Northern blot analysis for the effect of bovine decorin is shown in the left panel as described above. The reversion effect of skeletal muscle decorin on myogenin expression was evaluated using A6 myoblasts transiently cotransfected with pMyoLuc and pRL plasmids. The cells were incubated for 24 h in growth medium, harvested, and dual luciferase activity was determined as described under "Experimental Procedures." C, A6 cells were incubated with skeletal muscle decorin pretreated with chondroitinase ABC (Cabc) and assayed as explained in the legend of B. Decorin concentrations are as follows: D1 = 0.1 µM and D2 = 0.5 µM. The results shown are the means from two different experiments performed in duplicate.


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Fig. 6.   Concentration dependence of TGF-beta -mediated inhibition of myogenin expression in control and antisense decorin cells. Wild type (WT) and antisense decorin-transfected cells (A6) were incubated for 30 h in differentiation medium containing the indicated concentrations of TGF-beta 1 (A, left panel) and FGF-2 (B, left panel). RNA was isolated from the cells, and 10 µg of total RNA was analyzed by Northern blot with a 32P-labeled myogenin (Myo) cDNA probe. Right panels show the graphical representations of TGF-beta 1 (A)- and FGF-2 (B)-dependent inhibition of myogenin expression in antisense decorin cells (open circles) and wild type cells (closed circles). Values correspond to the means of three independent experiments. C shows TGF-beta 1-dependent inhibition of myogenin expression in wild type cells (closed circles) and wild type cells infected with adenovirus containing the full-length sequence for human decorin (open circles). Myogenin expression was evaluated as described in the legend of Fig. 4B.





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