Figure 5: Upstream and downstream mechanisms of mLLP acting on neural development.

(A) Neuronal activation by KCl treatment for 2 h at DIV 7 decreases mLLP protein expression level (right panel: quantification data of mLLP/GAPDH, n = 8 per group, unpaired t-test: **p = 0.0044). Loading controls (GAPDH) were detected on the same blot. Samples from the two groups were processed in the same experiment in parallel. Extended blot images including this data are presented in Supplementary Fig. 6B. (B) Nuclear extracts from HEK293T cells transfected with 3XFLAG-tagged mLLP or control plasmid were immunoprecipitated with anti-FLAG affinity gel and immunoblotted with antibodies against each protein indicated. All data are derived from the same blot, and the samples were processed in the same experiment in parallel. Extended blot images are presented in Supplementary Fig. 8. (C) qRT-PCR analysis of a subset of known CTCF target genes after mLLP knockdown in dissociated hippocampal neuron cultures at DIV 6 (unpaired t-test: **p < 0.01, *p < 0.05, n = 4 per group). All data are represented as mean ± SEM.