Suppression of TNF mRNA expression is accompanied by inhibition of apoptosis and subsequent milder demyelination in ibudilast-treated twi/twi at PND45. A, B, E, F, I, J: CWM, C, D, G, H, K, L: sp5. A-D: In situ hybridization of TNFα mRNA in vehicle-treated twi/twi (A, C) and ibudilast-treated twi/twi (B, D). Whereas vehicle-treated twi/twi show abundant signals in CWM (A) and sp5 (C), TNFα mRNA signals are remarkably reduced in the ibudilast-treated twi/twi (B, D). Inset in "A" shows TNF-α mRNA-positive microglia. E-H: TUNEL staining of vehicle-treated twi/twi (E, G) and ibudilast-treated twi/twi (F, H). Ibudilast-treated twi/twi shows fewer TUNEL-positive cells than are seen in vehicle-treated twi/twi. Arrowheads indicate TUNEL-positive cells. I-L: LFB-PAS staining of vehicle-treated twi/twi (I, K) and ibudilast-treated twi/twi (J, L). In the ibudilast-treated twi/twi, CWM and sp5 show much milder demyelination than in vehicle-treated twi/twi. Scale bar = 100 μm (I-L), 50 μm (A-H), 10 μm (inset in "A").