Figure 10.

An Akt inhibitor blocks collagen I protein expression in response to NoC1. MEFs cultured in media with 2 μmol/L ascorbic acid were treated daily for three days with either PI3-kinase inhibitor, 5 μmol/L LY294002 (LY), 2 μmol/L Akt inhibitor II (II), or DMSO vehicle (NT) in the presence or absence of 30 nmol/L NoC1. Cell layers harvested in Laemmli buffer were separated by SDS-PAGE and immunoblotted for type I collagen. Note that the processed and unprocessed procollagen bands were not resolved in the Laemmli extracts of the total cell lysates. Blots were stripped and reprobed for β-tubulin as a loading control. Blots were analyzed by densitometric analysis, and collagen levels were normalized to the loading control for each sample. Results are expressed as the mean fold change as compared to cells treated with DMSO ± SD (n = 3).