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. Author manuscript; available in PMC: 2022 Jun 6.
Published in final edited form as: Nature. 2022 Feb 14;602(7898):647–653. doi: 10.1038/s41586-022-04396-8

Fig. 3 |. Reduced oligodendrocyte maturation in 4EP+ mice.

Fig. 3 |

a, Number of differential genes in brain regions (n=5) of baseline mice; QuantSeq, one-way ANOVA Tukey test. b, Cell-specific enrichment analysis; p-values<0.05 in white. c, Representative subset of differential OPC and MO genes, one-way ANOVA Tukey with boxes showing 25–75th percentiles, median, and min/max points (n=5). d, Whole brain correlation to PVT seed using 2DG uptake data (n=11). PVT (red cross-section) is indicated. One-tailed multiple linear regression with minimum threshold of 200 contiguous voxels with p<0.05 was performed. e, Representative images of brain sections from 4EP- and 4EP+ mice; NG2 (red) and CC1 (green). Scale 100µm. f, Maturity quotient of oligodendrocytes in the PVT (n=8 cumulative totals used from biologically independent mice). p=0.004. g, Quantitative flow cytometry of PVT cells comparing the MOG+ quadrant 4 population over the NG2+ quadrant 1 population (n=4 pools of 5 PVTs). p=0.03. h, Representative flow plots. i-l, Organotypic brain slices treated with 10μM 4EPS. Each datapoint represents separate biological replicates. i, Maturity quotient of oligodendrocytes; using total cumulative counts from 3–5 images per replicate (vehicle, n=9; 4EPS, n=10). p=0.003. j, Percent colocalization of neurofilament (NF) and proteolipid protein (PLP) antibody stain (n=6). p=0.01. k, Representative images of panel j. Scale 20µm. l, Western blot for MBP of slices (n=6). p=0.04. Abbreviations: PVT, paraventricular nucleus of thalamus; BNST, bed nucleus of stria terminalis; BLA, basolateral amygdala; mPFC, medial prefrontal cortex; HY, hypothalamus; vHPC, ventral hippocampus; Astro, astrocyte; OPC, oligodendrocyte precursor cell; NFO, newly formed oligodendrocytes; MO, mature oligodendrocytes; Micro, microglia; Endo, endothelia; CC1, adenomatous polyposis coli; NG2, neural/glial antigen 2; MBP, myelin basic protein. Biologically independent mice from multiple randomized litters examined over one (a-c) or two respective experiments (d-l). Data represent mean ± SEM. Panels e-l analyzed by two-tailed Welch’s t-test. *p ≤ 0.05; **p ≤ 0.01.