Fig. 4

Knock-down of NetB rescues the MB defects and ameliorates the memory in dfmr1 mutants. a Knock-down of NetB rescued the β lobe fusion in dfmr1 mutants. b, c Western blot and RT-qPCR showed that NetB protein level was obviously elevated in the brains of dfmr1 mutants while mRNA level was not significantly altered compared with control. d, e Interaction of FMRP with NTN1 mRNA in HEK293 cells. d Western blot analysis showed FMRP was precipitated by the anti-FMRP antibody. IgG IPs were used as a negative controls. e RT-qPCR analysis indicated that NTN1 and MAP1B (positive control) mRNAs were significantly enriched in anti-FMRP IPs. ^*p < 0.05, ^**p < 0.01. f Polyribosome profile of NTN1 mRNA indicated a clear increase of the NTN1 mRNA in the most actively translating ployribosomes in the Fmr1 mutant lymphoblastoid cells compared with normal control (N = 3, **p < 0.01). g Knock-down of NetB in dfmr1 mutants ameliorated the courtship activity, immediate recall memory and short-term memory of dfmr1 mutant flies (^*p = 0.0328, ^**p = 0.0085, ^****p < 0.0001, Two-way ANOVA). h Knock-down of NetB in dfmr1 mutant flies by elav-Gal4 ameliorated the olfactory conditioning short-term memory of dfmr1 mutants (^*p < 0.05, ^***p < 0.001, One-way ANOVA). Error bars = SEM