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.