Figure 1. RT–PCR and immunohistochemistry demonstrate RhoA knockdown. A: Reverse transcription polymerase chain reaction (RT-PCR) shows a statistically significant (*p<0.05; ***p<0.0001) reduction in endogenous RhoA mRNA in eyes treated with increasing concentrations of siRhoA. The percentage immunoreactivity (IR) pixel counts of RhoA expression in the trabecular meshwork (TM) for the groups are defined in C–G. B: Note the statistically significant (*p<0.0001 difference from PBS/siRhoA) upregulation of RhoA protein in the TGF-β1-treated eyes and downregulation after knockdown with siRhoA. Radial rat eye sections immunohistochemically stained for RhoA (green) nuclear counterstaining with 4′,6-diamidino-2-phenylindole (DAPI) (blue) after intracameral (IC) injection with PBS for 35 days (C), TGF-β1 for 35 days (D), TGF-β1+siRhoA for 7 days (E) and 35 days (F), and TGF-β1+siEGFP for 35 days (G). Images were taken of the iridocorneal angle (C–G; *TM;^†Schlemm’s canal). Note the successful knockdown of RhoA in the TM of the IC siRhoA-treated eyes (E and F). All images are representative of the eight images taken per treatment group (scale bar=50 µm).