Figure 4. Autoradiograms of SDS-PAGE separated extracts from lenses incubated in radiolabelled amino acids

Fresh and 7 day cultured lenses were incubated for 5 h in medium containing 300 μCi ³⁵S-labeled amino acids. Lenses were carefully rinsed, weighed and homogenized in SDS-sample buffer. Protein (100 μg) from each sample was loaded onto NuPage 4-12% Bis-Tris gels. The left panel shows data for 4 fresh lenses and 4 cultured lenses. In the right panel selected regions are shown for 2 samples from each group at higher magnification. The bands indicated by numbers are among those with altered synthsis following 7 days of culture. The following numbered bands were identified by mass spectrometry. Band were identified as Filensin (1), Heat Shock Cognate 70 (2), Vimentin (3), Phakinin (4), βB1-crystallin (7). Bands 5 and 6 showed altered level of synthesis in the cultured lenses but have not been identified.