Figure 1: Subcellular localization and intrinsic disorder of mLLP protein.

(A) Nuclear/nucleolar localization of mLLP. Anti-fibrillarin (nucleolar marker) immunocytochemistry of neurons transfected with plasmid encoding mLLP-EGFP. Scale bar, 5 μm. (B) Diagram representing the various deletion mutants of mLLP fused to EGFP. mLLP full-length (FL) sequence, mutant constructs (ΔN or ΔC) absent of N-terminal (1–20) or C-terminal (107–130) parts, or either N- or C-terminal sequences were cloned into the pEGFP-N1 multiple cloning site. (C) Representative fluorescence microscopy images of neurons expressing EGFP fused with mLLP deletion mutants. The construct expressing mCherry driven by neuron-specific CaMKII promoter was co-transfected. Scale bar, 10 μm. (D) Quantitative measurement of the ratio of nuclear/cytosolic EGFP signals in (C). EGFP signal intensity normalized by that of mCherry was compared between in the nucleus and in cytosol of the somatic region. The dotted line indicates the value 1, which means no differential localization in the nucleus and cytosol. One sample t-test (hypothetical value 1), **p < 0.01, ***p < 0.001. Data are represented as mean ± standard error mean (SEM). (E) Disorder analysis of mLLP using two different methods VSL2B (blue) and PONDR-FIT (P-FIT, green) in DisProt. The residues with value exceeding 0.5 are considered disordered.