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. 2022 Jun 30;17(6):e0269404. doi: 10.1371/journal.pone.0269404

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© 2022 Kragness et al

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Fig 2 — A) Immunocytochemistry with HA antibody shows Pants localization throughout cultured hippocampal neurons from Pants^SOG-Y/+ mice, but not wildtype controls. B) Immunocytochemistry of 18–21 div primary neurons from Pants^SOG-Y hippocampus stained for HA, a Pre-synaptic marker (Synapsin 1a/b) and post-synaptic markers (MAP2 and Shank2) imaged with confocal microscopy. White arrows indicate areas of colocalization, and red arrows indicate suspected extra-synaptic pants that fails to co-localize with any markers. C) Structured Illumination Microscope imaging of the same staining from (A) to obtain better resolution. D) HA Western blot of HA IP from Nero2a cellular lysates (C), extracellular matrix (E) fractions, and proteins precipitated from cell media (M) after transfection with a construct for over-expressing the Pants^SOG-Y fusion protein compared to untransfected controls. E) HA Western blot of HA IP of media collected from primary Pants^SOG-Y hippocampal neurons or Pants^+/+ negative control neurons.