Fig. 4.

Gpi1 functions are modulated by Oxr1. a Over-expression of Gpi1, Oxr1-FL, and Oxr1-C in N2a cells treated with arsenite and quantification of pyknotic nuclei as a measure of cell death (N = 3 independent repeats). b Knockdown of Oxr1 and Gpi1 by shRNA in arsenite-treated N2a cells with quantification of pyknotic nuclei (N = 5 independent repeats). c Knockdown of Oxr1 and Gpi1 by shRNA together with either Gpi1 or Oxr1 over-expression, respectively, with quantification of the number of pyknotic nuclei (N > 5 independent repeats). d Wild-type CGCs cultured on Transwell inserts were treated with increasing concentrations of recombinant of Gpi1 and the number of cells having migrated through the insert were quantified (N = 2 repeats). e Representative images of CGC migration after 24-h treatment with recombinant Gpi1. Scale bar: 200 μm. f Quantification of migration coefficient in Oxr1^d/d and Oxr1^+/+ cultures (N > 3 animals per group). g–h Relative composition of primary cultures treated with either vehicle or recombinant Gpi1 and quantified by immunocytochemistry using a neuronal marker (NeuN) and a marker for proliferating cells (Ki67). Scale bar: 100 μm. i Expression of Gpi1 and its receptor, Gp78, in CGCs from Oxr1^d/d mice compared to Oxr1^+/+ by qRT-PCR (N = 4 animals per group). j Gp78 RNA expression levels in whole brain or cerebellum from P18 Oxr1^d/d mice compared to control littermates by qRT-PCR (N = 4–5 animals per group). Panels a–d: one-way ANOVA; f, h–j: t test; *p < 0.05, **p < 0.01, ***p < 0.001 as compared to shRNA scramble plus empty vector, shRNA scramble, empty vector, Oxr1^+/+ or vehicle; ^###p < 0.001 as compared to shRNA Oxr1 plus empty vector