Figure 9.

Effect of cadherin-reactive beads on myogenin expression in sparse cultures of C2 myoblasts. C2 cells were seeded and cultured overnight at 10% confluence, and then the medium was replaced with fresh FCS-containing “growth medium” (FS) or with “differentiation medium” containing horse serum and insulin (HI), and the various beads were added. (A) Total cell extracts were prepared at the indicated time points after addition of beads, subjected to SDS-PAGE, transferred to nitrocellulose sheets, and reacted with anti-myogenin antibodies. The absorbance of the myogenin bands, determined by densitometry, is shown. (B) The percentage of the myogenin positive nuclei after treatment was calculated. The cells were maintained in growth medium and treated with beads for 12 or 18 h, fixed, and immunolabeled for myogenin. The percentage of positive cells was calculated out of the entire population or the subpopulation of bead-associated cells. Every column represents the mean ± SD of 3 independent experiments, each in duplicate. In each individual experiment a total number of 900 cells or 200 bead-bound cells were counted. B, BSA; α, anti-N-cadherin antibodies; N, NEC.