Extended Data Fig. 1: GPR158 is present at the PC of hippocampal neurons.

A) PCA biplot showing that the proteomic datasets collected after stereotactic injections with either vehicle (NaCl) or OCN are orthogonal to each other. B) Bar graph representation of the top upregulated signaling pathways in differential analysis of hippocampi 4 h after stereotactic injections with either vehicle or OCN. C) Protein ontology network associated to PC-core proteins and PC functions enriched in the hippocampus after OCN treatment. D) Western blot analysis and quantification of LC3B-II accumulation in primary neurons infected with lentivirus expressing either shRNA-Gpr158 or shRNA-scramble. Independent neuronal cultures (n = 3) were treated with either vehicle, OCN, bafilomycin A1 (Baf), or OCN + Baf. E) Western blot analysis of LC3B-I, LC3B-II and IFT20 in WT and Gpr158-/- mouse hippocampi 4 h after hippocampal stereotactic injections with either vehicle or OCN. F) Schematic representation of the 5 discrete ciliary targeting (VXPX motif) sequences in the C-terminal tail of GPR158. We have generated a mutated Gpr158 version. G) Proportion of neuronal primary cilium presenting GPR158+ and ACIII+ puncta. H) Representative images of GPR158 staining in hippocampal cultures derived from either WT or Gpr158-/- mice. Scale bar=10μm. I-J) Representative images of co- immunofluorescence for GPR158 (green) and ACIII (orange) in hippocampal neurons from 3 independent experiments (I) and in WT hippocampal cross-section of the CA3 region of mice treated with OCN from 2 independent experiments (J). 1, 2, and 3 squares in I are 3D rendering images of GPR158 localization in PC from mature hippocampal neurons treated with vehicle or OCN. Scale bar=5μm. In all western blots, β-actin was used as a loading control for each sample. Data are expressed as mean ± SEM. The p values were determined by a one-tailed Student’s t-test.

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