Fig. 3
From: Rapid isolation of intact retinal astrocytes: a novel approach
RT-qPCR and immunohistochemistry of astrocytic enrichment from pull-off and pull-off plus immunoprecipitation in naïve mouse. A: RT-qPCR results showing fold-change enrichment of markers of astrocytes (GFAP), Müller cells (Cralbp), inner retinal neurons (Pax6), and outer retinal neurons (NeuroD1) in pull-off (PO) and pull-off plus immunoprecipitation (PO + IP), relative to whole retina control. B–E′: Immunostaining of pull-off isolated tissue shows colocalization of astrocytes (C) with HA (D). Although few non-astrocyte cells remain after the pull-off (B), the restriction of HA-tagged ribosomes to astrocytes enables further enrichment of astrocytic RNA during the ensuing immunoprecipitation. HA-tagged ribosomes are present throughout the processes of isolated astrocytes (E′). Scale bar in panels B–E = 100 µm and in E′ = 50 µm. F–I: Absence of Iba-1+ microglia on pull-off isolated tissue. Scale bar in all panels = 100 µm. J–M: Absence of Cralbp+ Müller cells on pull-off isolated tissue. Scale bar in all panels = 100 µm