Fig 2: N,N-dimethyltryptamine (DMT) promotes stem cell differentiation toward all neural phenotypes.SGZ-derived neurospheres were cultured for 7 days in the presence of DMT alone or in combination with the antagonists BD1063 (BD), methiothepin (Met), ritanserin (Rit), and WAY100635 (WAY) and then were adhered on a substrate and allowed to differentiate for 3 days. a Confocal fluorescent images showing the expression of the neuronal markers ß-III-Tubulin (TuJ-1 clone, green) and MAP-2 (red) in NS (n = 8 per condition). DAPI was used for nuclear staining. Scale bar = 50 µm. b Representative western blots and quantification of ß-tubulin and MAP-2. c Immunofluorescence images showing NS expressing the glial fibrillary acidic protein (GFAP, red) that stains astrocytes, and in green, the oligodendrocyte marker CNPase (n = 8 per condition). DAPI was used for nuclear staining. Scale bar = 50 µm. d Representative western blots of CNPase and GFAP and quantification. Results are the mean ± SD of four different cellular pools with three independent experiments/pool (n = 12). After confirming the significance of the primary findings using ANOVA, a significance level of P < 0.05 was applied to all remaining post hoc (Tukey test) statistical analyses. **P = 0.01; ***P = 0.001 indicate significant results versus non-treated (basal) cultures.

Fig 3: Oligodendrocyte lineage expression of LRF during the progression of demyelination and remyelination. Spinal cord transverse sections immunostained for LRF (red) in combination with CNP (A-C'; green) or Olig2 (D-G; green). In control white matter (A, D; PBS-injected match to 4 wpi MHV), LRF is present in the nuclei of oligodendrocytes labeled by CNP (arrows in A') or by Olig2 (arrows in D'). The majority of Olig2 cells express LRF in control white matter (G). During demyelination (4 wpi; B, B'), lesion areas have a loss of CNP immunoreactivity and reduction of Olig2 cells in white matter (G). In demyelinated lesions, a high density of LRF + cells are not labeled with CNP (B') or Olig2 (E'), indicating infiltration of non-oligodendrocyte lineage cells. As remyelination progresses, CNP immunolabeling shows myelin throughout the white matter although mild vacuolization and cellular infiltrate are still visible adjacent to the ventral root exit zone, a typical lesion site (8 wpi; C, C'). Many cells immunolabeled for CNP (arrows in C') or Olig2 (arrows in F') exhibit nuclear LRF at 8 wpi. Olig2 cell populations recover by 8 wpi and the distribution of Olig2 cells is almost evenly split among those with or without LRF (G). Olig2 nuclear immunoreactivity was used to identify oligodendrocytes, as was clearly visible even at lower magnifications (D-F). A low intensity of Olig2 immunoreactivity in processes was detectable in lesion areas at higher magnifications (E', F'), which was considered non-specific signal due to changes in the lesion tissue. One way ANOVA using n = 3 mice for each condition; p = 0.0022, F (3,8) = 12.46 for Olig2 + LRF- cells; p = 0.0009, F (3,8) = 16.29 for Olig2 + LRF + cells. Scale bars A-C, D-F = 200 um, A'-C' – 75 um, D'-F' = 25 um. VH = spinal cord ventral horn.