Silencing SMG1 expression increases p-syn and t-syn levels.

Shown in A are westerns confirming that SMG1 silencing using four unique siRNA sequences causes large increases in p-syn and t-syn, confirming screening results shown in Figure 2. Expression levels as a % of non-silencing (NS) siRNA control are under each western. Values are the average of all four siRNA sequences. siRNAs used are indicated across the top of the westerns. Antibodies used are anti-SMG1 (α-SMG1), anti-pS129 α-synuclein (α-pSyn), anti-α-synuclein (α-tSyn), and anti-α/β Tubulin (α-Tubulin). Shown in B are westerns demonstrating that SMG1 silencing increases expression of both endogenous α-synuclein (Tet+ samples) and transgenic α-synuclein that is expressed from the tet-off promotor (Tet- samples). Expression levels as a % of NS siRNA control are indicated beneath each sample. ND stands for not detected. Antibodies used are the same as in A. Results are representative of three independent replicates.