Fig 1.
Pedigrees of family 1 and family 2 with mutations in the UBAP1 gene.
Squares and circles indicate males and females, respectively. Black symbols represent members with an HSP phenotype and empty symbols represent unaffected individuals. Arrows indicate the probands.
Table 1.
Clinical features in HSP patients of the two families.
Table 2.
Primers used for screening mutation in UBAP1.
Fig 2.
Confirmation of novel variants of UBAP1.
(A) Sequence chromatogram showing the c.468_469delTG (family 1) and c.512T>G (family 2) variants in UBAP1 in the probands. The variants were numbered according to GenBank NM_016525.4. (B) Sequence alignment of mammalian UBAP1 protein showed that the regions around the variants were highly conserved. The numbers on the left and right indicate where the fragment was located within the entire protein sequence. The positions of the variants are marked by black triangles. (C) Mutations identified in SPG80. The numbers indicate the locations of mutations in DNA and protein. The mutations found in this study are marked in red.
Fig 3.
Characterization of A157* and L171* mutants.
(A) mRNA expression levels of UBAP1 in lymphocytes were analyzed by qPCR assays. Data were normalized to the levels of GAPDH within each sample (n = 3). The data are presented as the mean ± SE. (B) Protein levels of UBAP1 in lymphocytes were examined by Western blotting. GAPDH was used as an internal control. (C) Localizations of FLAG-tagged A157* and L171* analyzed in neuro-2a cells via immunofluorescent assays showing they did not disrupt the subcellular localization of UBAP1.
Fig 4.
A157* and L171* mutants disrupt neurite outgrowth in neuro-2a cells.
(A) Neuro-2a cells were transfected with the indicated constructs, and FLAG-positive cells were determined by immunofluorescent assays. (B) (C) The average length of each neurite and the numbers of neurites per cell were counted. One hundred neurites in FLAG-positive cells were scored for neurite outgrowth. Neurite numbers were counted in one hundred FLAG-positive cells. The data are presented as the mean ± SE (***p < 0.0001).