Cortical astrocytes from dual reporter mice were cultured alone or on top of an intact monolayer of bEND.3 cells and infected with an shRNA directed against RBPJ (shRBPJ), a scrambled shRNA control (shScr), or no virus 3–5 days after re-plating. Cell lysates were harvested for Western blot 11–13 days after transduction for analysis of (a) RBPJ, (b) eGFP, and (c) GLT-1 protein levels. Top: Representative blots and Bottom: summary of quantification, normalized to β-actin. Data are the mean ± SEM of six independent experiments. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001 for indicated comparisons.