Fig. 1.

shRNA-mediated knockdown of WT, P23H, T17M, and shRNA820-resistant (RHO₈₂₀) variants of human RHO. HEK293T cells were transfected with a plasmid expressing WT, P23H, T17M, or shRNA820-resistant (RHO₈₂₀) human RHO with a C-terminal turboGFP tag (RHO-tGFP) and with a rAAV2 plasmid (denoted in lane labels) encoding empty DNA (no shRNA), a control shRNA, shRNA₁₃₁, shRNA₁₃₄, or shRNA₈₂₀. A no-DNA transfection control was also included. (A, C, E, and G) Immunoblots of protein samples isolated from transfected HEK293T cells probed for turboGFP tag (green) and β-tubulin (red) as the loading control. Rho aggr., aggregated form of RHO-GFP; Rho mono., monomeric form of RHO-GFP. (B, D, F, and H) Relative quantification of the monomeric form of RHO-GFP (Upper) and of the monomeric and aggregated forms of RHO-GFP (Lower). The first lane of each Western blot contained the Chameleon Duo Prestained Protein Ladder from Li-Cor. Bars denote the mean value of three technical replicates; error bars denote SEM. ns, not significant, **P < 0.01, ***P < 0.001, ****P < 0.0001.