Figure 3.
Reduced SREBF2 and its downstream targets in the TDP-43–deleted oligodendrocytes. (A) LDLR and SREBF2 mRNA expression in oligodendrocytes of ctrl and cKO mouse lumbar spinal cord (arrowhead), revealed through combined RNA fluorescent in situ hybridization (RNA-FISH) and GST-P1 IF staining. Images were taken from the ventral gray matter at P21 and P60, at 20× magnification. Scale bar: 20 µm. 3D reconstruction of oligodendrocytes for LDLR (green) and SREBF2 (magenta) mRNA quantification. DAPI (blue). Scale bar: 3 µm. (B) Quantification of SREBF2 (i, ii) and LDLR (iii, iv) puncta in oligodendrocytes of ctrl and cKO mouse lumbar spinal cord, at P21 (i, iii) and P60 (ii, iv). Puncta counts for individual oligodendrocytes (faded circle and triangles), and means (solid circle and triangle) for each animal (n = 3) plotted, total mean ± SEM derived from all animals. Significance was tested using unpaired t test; *, P < 0.05; **, P < 0.01; ***, P < 0.001. n = 3 per genotype, 10 cells were quantified per section, and at least five sections were quantified per animal. (C) Confocal images of colabeling of oligodendrocytes (CC1+, red) and key proteins involved in cholesterol metabolism (SREBF2, left; HMGCS1, middle; and LDLR, right, green) in the white matter of spinal cord sections from 60-d-old mice. Square areas were separated into individual channels, indicating reduction of SREBF2, HMGCS1, and LDLR protein in oligodendrocytes. n = 3 per genotype, six to eight slices per animals were stained and observed. Scale bar: 10 µm. Enlarged images of single cell split into individual channels for SREBF2/HMGCS1/LDLR (green) and CC1 (red). Scale bar: 10 µm. cKO, conditional knockout; ctrl, control.

Reduced SREBF2 and its downstream targets in the TDP-43–deleted oligodendrocytes. (A) LDLR and SREBF2 mRNA expression in oligodendrocytes of ctrl and cKO mouse lumbar spinal cord (arrowhead), revealed through combined RNA fluorescent in situ hybridization (RNA-FISH) and GST-P1 IF staining. Images were taken from the ventral gray matter at P21 and P60, at 20× magnification. Scale bar: 20 µm. 3D reconstruction of oligodendrocytes for LDLR (green) and SREBF2 (magenta) mRNA quantification. DAPI (blue). Scale bar: 3 µm. (B) Quantification of SREBF2 (i, ii) and LDLR (iii, iv) puncta in oligodendrocytes of ctrl and cKO mouse lumbar spinal cord, at P21 (i, iii) and P60 (ii, iv). Puncta counts for individual oligodendrocytes (faded circle and triangles), and means (solid circle and triangle) for each animal (n = 3) plotted, total mean ± SEM derived from all animals. Significance was tested using unpaired t test; *, P < 0.05; **, P < 0.01; ***, P < 0.001. n = 3 per genotype, 10 cells were quantified per section, and at least five sections were quantified per animal. (C) Confocal images of colabeling of oligodendrocytes (CC1+, red) and key proteins involved in cholesterol metabolism (SREBF2, left; HMGCS1, middle; and LDLR, right, green) in the white matter of spinal cord sections from 60-d-old mice. Square areas were separated into individual channels, indicating reduction of SREBF2, HMGCS1, and LDLR protein in oligodendrocytes. n = 3 per genotype, six to eight slices per animals were stained and observed. Scale bar: 10 µm. Enlarged images of single cell split into individual channels for SREBF2/HMGCS1/LDLR (green) and CC1 (red). Scale bar: 10 µm. cKO, conditional knockout; ctrl, control.

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