Fig. 1

Screening of novel E3 ubiquitin ligase for SPAST using human protein microarray. A Schematic showing the screening of E3 ubiquitin ligase for SPAST-M1 through HuProt™ v3.1 protein microarray. B Schematic diagram of HuProt.™ protein microarray data analysis and filtering method involving SPAST protein. C Quantification of the relationship between SPAST and ubiquitin pathway-related genes using the array. D mRNA analysis of Spast-M1, Fbxl17, and neurogenesis marker genes from forebrain (FB) to spinal cord (SC) tissues of mouse embryos at indicated days. E Western blot analysis of SPAST-M1 or M85, FBXL17, and MAP2 as a neuronal marker from FB to SC tissues of mouse embryos at indicated days. Protein intensities quantified using densitometry and normalized to β actin expression. F Immunostaining of SPAST-M1 and FBXL17 on the brain and SC at the embryonic day 14.5. Scale bar: 100 μm. LV; lateral ventricular. G During neural differentiation, mRNA expression of SPAST-M1 and FBXL17 from ReNcell CX at the indicated days. H During neural differentiation, protein expression of SPAST-M1 or M87, FBXL17, MAP2, and Tau from ReNcell CX. The MAP2 and Tau expressions were used as neural markers. All experiments were performed in duplicate, data are expressed as the mean of two samples with standard deviation, and results are representative of two independent experiments. (*P < 0.05; ** P < 0.01)