Extended Data Fig. 6. Extended results of oligodendrocyte analysis implicating oligodendrocyte differences in 4EP+ mice in vivo and in vitro.

a, Extended quantitation of flow cytometry with MOG+/NG2+ ratio by quadrant in the cerebellum, cortex, and hypothalamus (n=4). b-c, Western blot analysis of MOG (p=0.03) and MBP (p=0.002) markers (respectively) of mature oligodendrocytes from the PVT brain region. PVT punches from two mice were pooled per data point in quantitative data (n=6 pooled samples each group). For gel source data, see Supplementary Information Figure 1. d-g, Organotypic brain slices were cultured in the presence of 10uM 4EPS. d, Representative images of CC1, NG2 and Olig2 staining. Two cohorts of mice were used, with each data point in Fig 3i representing quantified images from samples from individual mice. Scale bar 20µm. e, Example high-magnification image of axon in organotypic brain slices, stained with antibodies specific to NF (red), and PLP (green), with DAPI (blue), taken from image set used for quantification in 3i and Extended Data Fig 6d. Scale bar 8µm. f, qPCR results of oligodendrocyte genes, Cspg4 (p=0.04), Mog (p=0.03), and Mpb from organotypic brain slices (n=7). g, Western blot image of mature oligodendrocyte marker MBP (quantified in main Fig. 3l). For gel source data, see Supplementary Figure 1. Abbreviations: NG2, neural/glial antigen 2; CC1, antibody (anti-adenomatous polyposis coli (APC) clone) that binds mature oligodendrocyte marker; MOG, myelin oligodendrocyte glycoprotein; MBP, myelin basic protein. Two cohorts of mice from multiple litters were used for experiments in this figure. Data represent mean ± SEM. Statistics were performed using two-tailed Welch’s t-tests. * p ≤ 0.05; ** p ≤ 0.01.