FIG. 4.

Aquaporin expression varies with substrate stiffness. (A): Expression levels of aquaporin-1 (AQP1) were determined by qPCR in primary HREC cultured on soft (0.5 kPa) and stiff (50 kPa) hydrogels. Cells were harvested at short (10 days) and long (4 weeks) time points. (B) Aquaporin-1 protein levels were determined by Western blot analysis. Protein was isolated from HREC on soft and stiff hydrogels, cultured in parallel with cells used for qPCR analysis. Twenty micrograms of total protein was loaded per lane, and blots were probed with anti-AQP1 antibody (Millipore AB2219 @ 1:1000). GAPDH was used as a loading control.