Figure 1

CAG::MAML-DN inhibits the expression of the Hes5 Notch reporter. (a) Schematic representation of the electroporation paradigm with plasmids introduced ex vivo into E5 chick retinas, cultured for 2 days, dissociated into single cells and analyzed by flow cytometry. A schematic of the MAML dominant-negative construct is shown. (b) Confocal images of vertically sectioned E5 chick retinas co-electroporated with CAG::Nucßgal, Hes5::GFP Notch reporter, and with or without CAG::MAML-DN and then cultured for 2 days. The scale bar shown in the bottom right picture denotes 40 µm and applies to all images. All images are oriented with the scleral side of the retina at the top of the image. (c) Flow cytometry dot plots of dissociated chick retinal cells electroporated ex vivo at E5 with a co-electroporation control (CAG::iRFP), Hes5::GFP, and with or without CAG::MAML-DN and cultured for 2 days. The bar graph on the right shows the percentage of Hes5::GFP reporter-positive cells within the electroporated population. The Shapiro–Wilk normality test was used to confirm the normal distribution. ** signifies p < 0.01 with a two-tailed student’s t-test. Each point represents one biological replicate. The columns represent mean, and the error bars represent standard deviation.