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A) Recombination strategy for tamoxifen (tmx; cyan)–mediated replacement of the HRas gene (grey, flanked by loxP sites) with HRasG12V (purple) in PlpCreERt2;FR-HRasG12V (pHRsG/+) mice. B) 2-month-old (2MO) mice treated with tmx are subjected to the complex wheel (CW) test, as well as histological and DTI-MRI analyses, 1-16 weeks later. C) Cell populations are analyzed in seven regions throughout the anterior-posterior (I-IV) and lateral “B” - central “C” axes of the corpus callosum (CC), unless otherwise disclosed. D) Immunostaining of coronal sections showing recombinant cells (GFP+, green arrowheads), OLs (GSTpi+, purple arrowheads), and GFP+GSTpi+ recombinant OLs (orange arrowheads) in the CC of WT and pHRsG/+ mice. E) Bar graph showing that the percentage (normalized to dapi) of recombinant cells in WT and pHRsG/+ mice is not significantly different (unpaired Student’s t test; males P= 0.14 and females P= 0.24). Genotype/sex color code and “n” per group in E, G : male WT (green) and pHRsG/+ (orange); female WT (black) and pHRsG/+ (red). F) Immunostaining picture indicating recombinant cells (green arrowheads), microglia (IBA1+, blue arrowheads), and OPCs (PDGFRa+, orange arrowheads) in the CC of WT and pHRsG/+ mice. G) The percentage of IBA+ cells in WT and pHRsG/+ mice (% of age/gender matched WTs) indicates significantly decreased # of microglia in male pHRsG/+ mice (unpaired Student’s t test; P= 0.047). Insets in D and F show the CC region in high magnification (dotted yellow square). Dapi was used to stain nuclei and normalize cell densities per arbitrary units (AU). Scale bar = 25μm. *P < 0.05.

Journal: bioRxiv

Article Title: Targeting Nitric Oxide Synthase 2 Reverses Learning Deficits in an Oligodendrocyte-Focused Model of Costello Syndrome

doi: 10.64898/2026.06.01.729333

Figure Lengend Snippet: A) Recombination strategy for tamoxifen (tmx; cyan)–mediated replacement of the HRas gene (grey, flanked by loxP sites) with HRasG12V (purple) in PlpCreERt2;FR-HRasG12V (pHRsG/+) mice. B) 2-month-old (2MO) mice treated with tmx are subjected to the complex wheel (CW) test, as well as histological and DTI-MRI analyses, 1-16 weeks later. C) Cell populations are analyzed in seven regions throughout the anterior-posterior (I-IV) and lateral “B” - central “C” axes of the corpus callosum (CC), unless otherwise disclosed. D) Immunostaining of coronal sections showing recombinant cells (GFP+, green arrowheads), OLs (GSTpi+, purple arrowheads), and GFP+GSTpi+ recombinant OLs (orange arrowheads) in the CC of WT and pHRsG/+ mice. E) Bar graph showing that the percentage (normalized to dapi) of recombinant cells in WT and pHRsG/+ mice is not significantly different (unpaired Student’s t test; males P= 0.14 and females P= 0.24). Genotype/sex color code and “n” per group in E, G : male WT (green) and pHRsG/+ (orange); female WT (black) and pHRsG/+ (red). F) Immunostaining picture indicating recombinant cells (green arrowheads), microglia (IBA1+, blue arrowheads), and OPCs (PDGFRa+, orange arrowheads) in the CC of WT and pHRsG/+ mice. G) The percentage of IBA+ cells in WT and pHRsG/+ mice (% of age/gender matched WTs) indicates significantly decreased # of microglia in male pHRsG/+ mice (unpaired Student’s t test; P= 0.047). Insets in D and F show the CC region in high magnification (dotted yellow square). Dapi was used to stain nuclei and normalize cell densities per arbitrary units (AU). Scale bar = 25μm. *P < 0.05.

Article Snippet: Floating sections were processed for immunodetection using antibodies for the reporter gene GFP (Nacalai Tesque, Kyoto, Japan), and the cell-type markers GSTpi (MBL Ltd, Tokio, Japan), CC1 (Calbiochem, San Diego, CA, USA), NG2 (Millipore, Burlington, MA, USA), PDGFRa (Rn’D systems, Minneapolis, MN), GFAP (Dako, Santa Clara, CA, USA), IBA1 (Wako, Osaka, Japan), and NeuN (Millipore, St. Louis, MO).

Techniques: Immunostaining, Recombinant, Staining