Figure 12. Hsp90 protein expression four days after treatment

A total of 12 lens epithelial explants from each of the eight treatment groups were used for western blot analysis of Hsp90 (90 kDa) protein expression. HeLa cell lysate (positive control), control (lanes 1 and 2), FGF-2 (lanes 3 and 4), TGF-β2 (lanes 5 and 6), and TGF-β2/FGF-2 (lanes 7 and 8) lens epithelial explants under normal conditions (lanes 1, 3, 5, and 7) and after heat shocked treatment (lanes 2, 4, 6, and 8) are shown. β-Actin (42 kDa) protein expression serves as an internal control and is used to normalize the protein band intensity. The bar graph represents the Hsp90 protein expression (percent of normal control; ±SEM) for control, FGF-2, TGF-β2, and TGF-β2/FGF-2 treated lens epithelial explant extracts from normal culture and heat shocked conditions. Statistical analysis (ANOVA: p is less than or equal to 0.05) shows that there is a significant treatment effect. The heat shocked control (55.6%±1.1%) and heat shocked TGF-β2-treated explants (73.6%±6.0%) demonstrate significantly lower levels of Hsp90 expression when compared to normal control (100%) epithelial explants. Although there is also a decrease of Hsp90 expression in the normal TGF-β2 (93.9%±2.5%), it is not significant (marked with a "+" sign). The FGF-2 (normal, 172.6%±2.5%; heat shocked, 129.0%±3.3%) and TGF-β2/FGF-2 (normal, 187.0%±5.3%; heat shocked, 157.0%±5.3%) explants show a significant increase of Hsp90 protein expression with the greatest increase in the TGF-β2/FGF-2 explants (marked with a black dot). An asterisk indicates that the Hsp90 expression of heat shocked FGF-2, TGF-β2, and TGF-β2/FGF-2 explants are significantly lower than the same treatment groups under normal culture conditions.