Functional Role of SIL1 in Neurodevelopment and Learning

Figure 5

Reelin signaling was impaired after SIL1 silencing. (a) Interactions between SIL1 and PSD95, VLDLR, and ApoER2 were examined by coimmunoprecipitation. Anti-SIL1, anti-PSD95, anti-VLDLR, and anti-ApoER2 antibodies were used to coprecipitate SIL1. GAPDH was used as a negative control. (b) The membrane fraction and total expression of VLDLR and ApoER2 were examined after incubation of neurons with silencing shRNA (shRNA#1 and shRNA#2) or scramble shRNA (con). (c) Statistical analysis of (b): the expression of each protein was first normalized to GAPDH and then compared with the control; both total expression and surface fraction of ApoER2 and VLDLR were significantly reduced (ApoER2 total: after shRNA#1 treatment, after shRNA#2 treatment; ApoER2 surface: after shRNA#1 treatment, after shRNA#2 treatment; VLDLR total: after shRNA#1 treatment, after shRNA#2 treatment; and VLDLR surface: after shRNA#1 treatment, after shRNA#2 treatment). All data were shown as . ^##. . (d) The phosphorylation of Dab1 after SIL1 silencing (shRNA#1 and shRNA#2) was assessed by antibody targeting phosphorylated Dab1. Neurons treated with scramble shRNA were used as the control (con). (e) Statistical analysis of (d): the phosphorylated Dab1 was first normalized to GAPDH and then to Dab1 and compared with the control (p-Dab1: after shRNA#1 treatment, after shRNA#2 treatment). All data were shown as . The phosphorylation of Dab1 was significantly inhibited by SIL1 silencing. ^##. for all treatments.

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