Figure 9. AND-34 expression regulates p130Cas and Akt signaling in lens epithelial cells. Lenses were removed from the eyes of AND-34^−/− and AND-34^+/+ mice (four months old), and the capsular epithelial layer (EPI) was separated from the lens fibers (FIB) by dissection. A: The expression of AND-34/NSP2 and CHAT/NSP3 were examined in the lens tissues of AND-34^+/+ mice by immunoblot. Lysates from HEK293T cells that were transfected with AND-34 (pAND) or CHAT (pCHAT) expression constructs were used as controls. Expression of crystallin, a component of lens fibers, and tubulin were also examined. To detect low level expression of AND-34 in lens fibers, fivefold more of the lens fiber sample was loaded in the first lane (Fib-5X) than in the fifth lane (Fib-1X). B: The phosphorylation status of p130Cas and AKT Ser 473 was examined in AND-34^−/− and AND-34^+/+ lens epithelial and lens fiber samples by western blot analysis. Phosphorylated p130Cas is characterized by reduced PAGE migration. Total levels of AKT expression were also assessed as a control. Similar results were obtained in three experiments.