Extended Data Fig. 2: Generation of iPKR mouse. | Nature Neuroscience

Extended Data Fig. 2: Generation of iPKR mouse.

From: Cell-type-specific drug-inducible protein synthesis inhibition demonstrates that memory consolidation requires rapid neuronal translation

Extended Data Fig. 2

a) Schematic showing the subcloning and targeting strategy of the multicistronic cassette containing loxP-flanked STOP cassette, NS3/4 protease, EGFP-L10, and iPKR kinase domain, which were separated by 2A ribosome skipping sites. The entire cassette was inserted between exon1 and exon 2 of Eef1a1 genomic locus in mouse ES cells. Recognition site for the Southern blot probe is indicated. b) Southern blot after BamHI restriction enzyme-digested DNA isolated from embryonic stem cells using the probe indicated in a). Modified (6.2 kB kb) and unmodified (10.3 kB) DNA bands are indicated with arrows. In Nes iPKR brains, EGFP-L10 is expressed in the soma of neurons in the anterior cingulate cortex (c), somatosensory cortex (d), CA1 (e), CA3 (f) and dentate gyrus (g) consistent with NeuN expression. Insets show the corresponding brain areas at higher magnification.

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