At 0 h PCS, only low levels of fibrotic ECM proteins (collagen I and
tenascin C) and the enzymatic ECM crosslinker Lysyl oxidase (Lox) are detected
in both WT and FNcKO LCs. Both WT and FNcKO LCs significantly upregulate
collagen I protein expression by 48 h PCS (WT ***P ≤
0.001; FNcKO ***P ≤ 0.001) although FNcKO LCs exhibit
less association with collagen I fibrils compared to WT (**P
≤ 0.009). By 5 days PCS, WT LCs expressing αSMA are associated
with a robust matrix of collagen I (***P ≤ 0.001) while
this is absent in the area surrounding FNcKO lens cells (WT vs FNcKO
***P ≤ 0.001). Similarly, Lox protein upregulates in
both WT and FNcKO LCs at 48 h PCS (WT **P = 0.004; FNcKO
*P = 0.020) while this signal is much less pronounced in
FNcKO LCs compared to WT (***P ≤ 0.001). There is a
significant increase in tenascin C fibrils surrounding WT LCs both at 48 h
(**P = 0.005) and 5 days (***P ≤
0.001) PCS compared to 0 h PCS. However, FNcKO lens cells are associated with
significantly less tenascin C fibrils both at 48 h PCS (***P
≤ 0.001) and 5 days PCS (***P ≤ 0.001) compared
to WT. Collagen I, Tenascin C, and Lox (red), αSMA (green), are merged
with DNA detected by Draq5 (blue). Scale bars: 35 μm. LC, remnant lens
epithelial cells/lens cells; C, lens capsule. All experiments had N = 3. Values
are expressed as mean ± SEM. Asterisks (*) indicate statistically
significant MFI between WT and FNcKO at a PCS or between two PCS time
points.