Fig. 3: PACS1R203W causes Golgi outposts to deploy into dendrites of hippocampal neurons. | Nature Communications

Fig. 3: PACS1R203W causes Golgi outposts to deploy into dendrites of hippocampal neurons.

From: Neural deficits in a mouse model of PACS1 syndrome are corrected with PACS1- or HDAC6-targeting therapy

Fig. 3: PACS1R203W causes Golgi outposts to deploy into dendrites of hippocampal neurons.The alternative text for this image may have been generated using AI.

a, b Western blots (a) and immunohistochemistry (IHC) of coronal brain sections (b) prepared from adult R26P1 and R26P1R203W mice induced or not with Emx1Cre to induce expression of HA-tagged PACS1 or PACS1R203W (red). Scale bar, 1000 μm. c Hippocampal CA1 region from specimens in (b). SO, stratum oriens; SP, stratum pyramidale; SR, stratum radiatum. Scale bar, 50 μm. d Dissociated Emx1Cre;R26P1 or Emx1Cre;R26P1R203W hippocampal neurons (DIV5) were fixed and stained for β3-tubulin (white, insets), Giantin, and HA-tagged PACS1 or PACS1R203W. High-resolution tiled images of entire cultured hippocampal neurons were captured using a confocal microscope (see Methods and Fig. S3c). (top) 3D surface reconstructions (Imaris, see Methods) of the captured images depict Golgi elements (red), HA-tagged PACS1 or PACS1R203W (green), and nuclei (blue). Scale bar = 5 μm. (Bottom, left and right) Higher magnification of R26P1 and R26P1R203W neurites (yellow arrowheads in insets in the top panels) containing HA-tagged PACS1 or PACS1R203W and Golgi (R26P1R203W neurons). Scale bar = 1 μm. (Bottom middle) Quantification of deployed Golgi elements based on their subcellular localization. Somatic Golgi, within cell body; Neurite Golgi, Golgi deployed into developing neurites. Data mean ± SEM (2-tailed Mann–Whitney U-test), n = 8 cells/group from two independent experiments. See also Supplementary Movie 1. Source data are presented as Sources Data files.

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