Figure 3.

Collagen I–induced N-cadherin up-regulation in NMuMG/E10 cells. (A) NMuMG/E10 cells were cultured on noncoated (non), collagen I-coated (col), or fibronectin-coated (fib) dishes. Two days after seeding cells were extracted and 30 μg of protein resolved by SDS-PAGE and immunoblotted for fibronectin, N-cadherin, and E-cadherin. (B) Immunoblots were quantified by densitometry using Adobe Photoshop and normalized to GAPDH. The columns represent mean values of three independent experiments and the bars represent the SD (∗p < 0.05; col versus non or fib). (C) RT-PCR was done for N-cadherin, E-cadherin, and GAPDH (as a control). (D) N-cadherin and E-cadherin mRNA levels were analyzed by quantitative real-time PCR. 18S rRNA was used as an endogenous control, and quantification of mRNA levels was performed in duplicate and repeated three times. The columns represent mean values and the bars represent the SD (∗p < 0.05; col versus non or fib). (E) N-cadherin mRNA was analyzed by quantitative real-time PCR from 3- to 48-h time points. Quantification was performed in duplicate and repeated two times.