Figure 1: Levels of Notch1 and Jagged1 in postnatal day 2 WT and Nuc1 optic nerve astrocytes.

(a,b) qRT–PCR using Taqman expression probes shows that Notch1 was elevated by >2-fold in Nuc1 astrocytes (*P=0.027), relative to WT astrocytes. WT and Nuc1 astrocytes had similar levels of Jagged1. (c) Western analysis for Jagged1 shows similar protein levels in WT and Nuc1 astrocytes. (d) Retinal flat mounts from a 3-day-old WT rat showing significant co-localization (yellow) of Jagged1 (red) with GFAP-positive astrocytes (green). (e,f) Immunostaining and western analysis for active Notch (NICD) using anti-V1744 antibody shows that NICD was decreased by 76% in Nuc1 astrocytes compared with WT astrocytes (*P=0.0031). (g) Expression and distribution of NICD and Jagged1 as revealed by immunostaining with anti-V1744 active Notch (red) and anti-Jagged1 antibodies (green). Decreased nuclear staining for NICD was evident in Nuc1 cells, but Jagged1 staining was similar in WT and Nuc1 astrocytes. Scale bar, 20 μm. In all panels, graphs show mean values and error bars represent s.d. from a triplicate experiment representative of at least three independent experiments. Statistical analysis was performed by a two-tailed Student’s t-test: *P<0.05.