Figure 4. Thrombin induces a molecular weight shift in extracellular Cyr61 at all time points and with increasing concentrations of
thrombin in human corneal stromal fibroblasts and myofibroblasts. Fibroblasts (
A,
C) and myofibroblasts (
B,
D) were treated with a serum-free medium alone (-) or containing 1.0 U/ml thrombin (+) from 1-7 h (
A,
B), or with 0 to 10.0 U/ml thrombin for 3 h (
C,
D). Conditioned media and cell lysates were collected, processed, and evaluated via western blot analysis for Cyr61 using an
antibody against the central linker region of Cyr61 (
Figure 1D). Samples were normalized to GAPDH (
A,
B) or total cell protein (
C,
D). Representative paired blots are shown. Arrowheads denote non-specific bands detected on secondary antibody only blots;
n = 2 independent experiments with different cell donors.