Figure 2.

Loss of Tsc1 in myelinating glia affects the myelination phase leading to axonal hypomyelination. (A,B) Immunostaining of cross cryosections of sciatic nerves from P15 Tsc1^Flox (A) and Tsc1^cKO (B) mice with antibodies against myelin basic protein (MBP; green; A,A’,B,B’) and pan-Neurofilament (NFL, red, A’,B’). White arrows indicate axons lacking MBP immunostaining surrounding axonal NFL (red). (C,D) Immunostaining of differentiated oligodendrocyte progenitor cell (OPC) cultures derived from P15 Tsc1^Flox (C,C’,C”,C”’) and Tsc1^cKO (D,D’,D”,D”’) mice, with antibodies again MBP (green) and Cnpase (red). DAPI was used as the labeling for nuclei. (E,F) Western blot analysis of MBP, myelin-associated glycoprotein (MAG) and PDGFRa proteins in SC (E) and MBP, MAG and P0 in sciatic nerves (F) from P15 Tsc1^Flox and Tsc1^cKO mice. β-actin was used as loading control. (G) Quantification of immunoblots shown in (E,F), *p < 0.05, or **p < 0.01. (H) RT-qPCR analysis of key differentiation genes in SCs from P15 Tsc1^Flox (green) and Tsc1^cKO (red) mice. Comparisons between genotypes were performed by Student t-test. n = 5, *p < 0.5. Scale bar: (A–E) = 2 μm.