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. 2019 Apr 30;116(20):10130–10139. doi: 10.1073/pnas.1821306116

Fig. 2.

Fig. 2.

Isolation and validation of oligodendrocyte-specific mRNAs from Olig1-RiboTag mice. (A) Overview of experimental strategy to isolate oligodendrocyte-specific RNAs from corpus callosum of Olig1-RiboTag mice. (B) Representative images from corpus callosum of Olig1-RiboTag mice validating the specificity of HA expression in oligodendrocytes. Immunofluorescence staining for HA showed abundant colocalization with the immature/mature oligodendrocyte marker CC1, as well as some colocalization with the mature marker GSTπ. No colocalization was shown with the astrocyte marker GFAP, neuronal marker NF200, and microglial marker Iba1. (Scale bar, 20 μm.) (C) Comparing HA-IP RNA samples vs. input RNA samples from corpus callosum (CC) of Olig1-RiboTag mice, qPCR showed enrichment of oligodendrocyte genes (Plp, Ugt8a, and Pdgfrα) and deenrichment of astrocyte (Aldh1l1), neuronal (Syp), and microglial (Tmem119) genes. (D) Representative images from optic nerve of Olig1-RiboTag mice validating the specificity of HA expression in oligodendrocytes in optic nerve. (Scale bar, 20 μm.) (E) Comparing HA-IP RNA samples vs. input RNA samples from optic nerve of Olig1-RiboTag mice, qPCR showed enrichment of oligodendrocyte genes (Plp, Ugt8a, and Pdgfrα) and deenrichment of astrocyte (Aldh1l1), neuronal (Gap43), and microglial (Aif1) genes.