BacTRAP-based ribosome profiling of murine hypothalamic PNOC neurons

We performed a BacTRAP-based ribosome profiling of PNOC-expressing cells in the hypothalamus of mice. Therefore, we employed mice, which express a fusion protein of the ribosomal L10a protein with EGFP (L10a-EGFP) under control of the PNOC promoter (Doyle et al., 2008). Precipitation of ribosomes of hypothalamic PNOC neurons with anti-GFP antibodies and subsequent mRNA sequencing of associated mRNAs allowed for assessment of an in-depth translational profile of these cells. Affinity purification of translating ribosomes was performed as described by (Heiman et al., 2014) with minor modifications. Overall design: Hypothalamic PNOC neuronal translatome of 10-12 week old PNOC-L10a-EGFP mice, fed either a normal chow diet (CD) for 3 days or a high-fat diet (HFD) for 3 days.