Fig. 7: Hx induces nuclear localization of Sirt2 in OPCs.
From: Sirt2 promotes white matter oligodendrogenesis during development and in models of neonatal hypoxia
a, c Coronal sections of subcortical WM from Nx and Hx WT mice at P18. Dotted lines delineate WM, WM-white matter. Scale bar = 100 µm. b Quantification of cytoplasmic and nuclear Sirt2 expression in PDGFRα+ OPCs (Nx vs Hx: ****p < 0.0001, *p = 0.0152, Student’s t test) d Quantification of cytoplasmic and nuclear Sirt2 expression in CC1+ OLs (Nx vs Hx: **p = 0.0042, ****p < 0.0001, Student’s t test). Graphs display mean ± SEM values (n = 4 brains per condition). e Experimental procedure for Sirt2 ChIP-seq using dissected subcortical white matter (SCWM) from P15 WT mice reared under Nx or Hx conditions. f The number of enriched Sirt2 binding peaks identified following comparison of Nx, Hx, and IgG (negative control) samples. g The location of Sirt2 peaks enriched after Hx. h Sirt2 ChIP-seq data were compared with previously published RNA-seq data from Hx OPCs39. Four genes that have nearby Sirt2 binding peaks are also altered in their expression following Hx. i QPCR verification of enriched Sirt2 binding at peaks 268-3, 268-2, and 308 in Hx WM, compared to Nx (**p = 0.0017, *p = 0.014, #p = 0.059 Student’s t test, n = 4 Nx and Hx samples). j Visualization of Sirt2 binding in genomic region upstream of Diaph2 gene. Red box highlights enriched binding in Hx. k RNAscope analysis of Diaph2 expression in Pdgfrα+ OPCs (arrows) in the WM of Nx and Hx WT mice at P22. Scale bars = 10 µm. l Quantification of the percentage of Diaph2+ OPCs in the WM of Nx and Hx WT mice at P22 (***p = 0.0001, n = 4 Nx and Hx mice, Student’s t test). m Visualization of Sirt2 binding in the genomic region upstream of Vegfc gene. Red box highlights enriched binding in Hx. n RNAscope analysis of Vegfc expression in Pdgfrα+ OPCs (arrows) in the WM of Nx and Hx WT mice at P22. Scale bars = 10 µm. o Quantification of the percentage of Vegfc+ OPCs in the WM of Nx and Hx WT mice at P22 (****p < 0.0001, Student’s t test, n = 4 Nx and Hx mice). Graphs display mean ± SEM values. All statistical tests are two-sided. Source data are provided as a Source Data file. The schematic in e was created using CorelDraw 2018 software (version 20.1.0.708).
