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. Author manuscript; available in PMC: 2014 May 19.
Published in final edited form as: Nat Cell Biol. 2013 May 5;15(6):614–624. doi: 10.1038/ncb2735

Figure 4.

Figure 4

SIRT1 inactivation does not alter the differentiation and myelination potential of oligodendrocytes. (a) Markers for specific cell types during differentiation of NSCs into mature oligodendrocytes. (b) Absence of active SIRT1 increases the percentage of O4-positive cells in differentiated adult NSC and neural progenitor cultures. Primary NSC and neural progenitor cultures from Sirt1lox/lox and NestinCre;Sirt1lox/lox 8-week-old mice were differentiated by growth factor removal and addition of 1% fetal bovine serum (FBS) after the first passage and immunostained after 2, 4 and 7 days for the oligodendrocyte marker O4. Images are representative of O4-positive cells at day 7 of differentiation. Arrowheads indicate individual O4-positive cells. Mean±s.e.m. of 5 independent experiments. Two-way ANOVA analysis: both genotype and differentiation day are significantly different, **P = 0.0055 and P = 0.0029, respectively. No interaction. Bonferroni post-test: day 7, P < 0.05. (c) SIRT1 inhibition enhances production of oligodendrocytes from NSCs and neural progenitor cells. Proliferating NSCs and neural progenitors were incubated with the SIRT1-specific inhibitor EX-527 (5 μM) or dimethylsulphoxide (DMSO) vehicle for 24 h. Cells were then switched to differentiation media (1% FBS) containing the same concentration of compound or vehicle and grown for a further 48 h. Cells were allowed to differentiate without added compound for 5 more days before staining for the oligodendrocyte marker O4. Mean±s.e.m. of 3 independent experiments. One-tailed, unpaired, Student’s t-test, *P < 0.05. (d) Absence of active SIRT1 increases the percentage of MBP-positive cells in differentiated adult NSC and neural progenitor cultures. Primary NSC and neural progenitor cultures from Sirt1lox/lox and NestinCre;Sirt1lox/lox 8-week-old mice were differentiated for 7 days by growth factor removal and addition of 1% FBS after the first passage and immunostained 7 days for the mature oligodendrocyte marker MBP. Images are representative of MBP-positive cells at day 7 of differentiation. Mean±s.e.m. of 6 independent experiments. Two-tailed, unpaired Student’s t-test, *P = 0.0398. Scale bars, 50 μm (b,d).