(A) NRG enrichment in P2 and synaptosomes in ctoNrg1 forebrains. Subcellular fractions were subjected to western blotting with indicated antibodies. Left, representative blots; right, quantitative data where NRG1 levels were normalized by synaptagmin1 (Syt1), and those in controls were taken as 1. n = 3 per genotype; *** Genotype F (1, 8) = 124.48, p < 0.001, two-way ANOVA. N, nucleus; C, cytoplasm; Syn, synaptosomes.
(B) Similar levels of LIMK1 in control and ctoNrg1 forebrains. Homogenates of individual control (lanes 1-4) and ctoNrg1 (5-8) mice were subjected to western blotting with antibodies against LIMK1 and α-tubulin. Left, representative blots; right, quantitative data where LIMK1 was normalized by α-tubulin. n = 4 per genotype; p > 0.05, t-test.
(C) Increased LIMK1 in P2 and synaptosomes of ctoNrg1 forebrains. Left, representative blots; right, quantitative data where LIMK1 in cytoplasm and P2 or synaptosomes were normalized by tubulin and Syt1, respectively. Levels in control mice were taken as 1. n = 3 per genotype; ** Genotype F (1, 12) = 17.73, p < 0.01, two-way ANOVA.
(D) Increased p-cofilin in P2 and synaptosomes of ctoNrg1 mice. Left, representative blots; right, quantitative data where p-cofilin was normalized by total cofilin. Data from controls were taken as 1. n = 3 per genotype; *** Genotype F (1, 8) = 26.68, p < 0.001, two-way ANOVA.
(E) Similar levels of LIMK1 and p-cofilin in P2 and Syn in aDox-treated control and ctoNrg1 forebrains. Left, representative blots; right, quantitative data where LIMK1 in cytoplasm was normalized by tubulin and that in P2 and synaptosomes by Syt1, p-cofilin was normalized by total cofilin; and those in controls were taken as 1; n = 3 per genotype; For LIMK1, Genotype F (1, 8) = 0.26, p > 0.05, two-way ANOVA; for p-cofilin, Genotype F (1, 8) = 0.00, p > 0.05, two-way ANOVA.
(F) HA-NRG1 was not detectable in forebrains of aDox-treated mice. Left, representative blots; right, quantitative data where NRG1 levels were normalized by Syt1, and those in controls were taken as 1; n = 3 per genotype; Genotype F (1, 8) = 1.57, p > 0.05, two-way ANOVA.