Figure 3: YAP directly regulates myelin and myelin-related protein expression

(a) Western blots of differentiated primary rat Schwann cells following infection with GFP control or constitutively active YAP S127A Flag adenovirus. GFP and Flag panels confirm infection in four independent cultures of each condition. Myelin proteins EGR2 (∼50 kDa) expression is increased, MPZ (∼27 kDa) is downregulated and MBP (∼45 kDa) is unchanged by YAP activation. A positive control for MBP from whole sciatic nerve extract is shown in insert, right. MAG (∼100 kDa) is also upregulated as is expression of proteins with relevance to myelination, Rab11a (∼20 kDa) and Laminin1 (∼175 kDa), in response to YAP activation. Actin (∼48 kDa) was used to verify loading. (b) Quantification and statistical analysis of the change in selected protein expression in primary RSCs following differentiation and infection with dominant active YAP S127A, relative to levels in cells infected with GFP control (normalized to 1, dotted line). (c) HEK 293T cells were transfected with plasmids expressing Mpz or Egr2 promoter driving Firefly luciferase expression, and either GFP, YAP alone or YAP and TEAD isoforms together. Luciferase levels were normalized to cells transfected with control GFP (three independent experiments in triplicates). Statistical analysis compares each transfection with the GFP infection for each promoter tested. a.u., arbitrary units. Statistical two-tailed Student’s t-tests: *P<0.05; **P<0.01; ***P<0.001; NS, P>0.05 not significant (NS). Error bars show s.e.m.