Figure 3: Sirt1 knockdown reduces proliferation of cultured OPCs after HX.

NX and HX white matter (P18) cells were transfected with Sirt1 siRNA and scrambled control, and labelled with BrdU (a) and NG2 (d). Scale bar, 100 μm. (b) Representative western blot analysis demonstrates Sirt1 knockdown in transfected NX and HX cells, compared with cells transfected with scrambled control (n=3 brains per group, per treatment). Sirt1 siRNA treatment shows a reduction in the percentage of BrdU⁺ cells (c) and NG2⁺ cells (e) in both NX (basal proliferation) and HX compared with controls. (f) Inhibition of Sirt1 activity by sirtinol diminished overall cell proliferation after HX (five NX and six HX brains for each treatment; four cultures for each conditions; *P<0.05; **P<0.01; ****P<0.0001, one-way analysis of variance, Bonferroni post hoc test, mean±s.e.m.).