Figure 1: Characterization of pSuper-mRbfox1-iso1 vectors. | Scientific Reports

Figure 1: Characterization of pSuper-mRbfox1-iso1 vectors.

From: Essential role of the nuclear isoform of RBFOX1, a candidate gene for autism spectrum disorders, in the brain development

Figure 1

(a) Knockdown of exogenous Rbfox1-iso1 in COS7. pCS-MT-mRbfox1-iso1 (Myc-iso1) was cotransfected into COS7 cells with pSuper-H1.shLuc (Control), pSuper-mRbfox1-iso1#1 or -iso#2. After 48 h, cells were harvested and subjected to western blotting with anti-Myc (upper panel). Anti-Sept11 was used for loading control (lower panel). Relative band intensity was also shown. (b) Knockdown of endogenous Rbfox1-iso1 in neurons. pCAG-EGFP was transfected with pSuper-H1.shLuc (Control) or pSuper-mRbfox1-iso1#1 into dissociated mouse hippocampal neurons obtained at E16, and cultured in vitro for 72 h. After fixation, cells were immunostained with anti-GFP (chicken; green) and anti-Rbfox1 (magenta). Scale bar shows 10 μm. Quantification of Rbfox1 expression was performed with ImageJ by analyzing the nuclei of control and deficient cells (arrowheads). (c) Effects of mRbfox1-iso1-knockdown on expression of Rbfox2 and Rbfox3. pCAG-Myc-mRbfox2 or –mRbfox3 was cotransfected into COS7 cells with pSuper-H1.shLuc (Control), pSuper-mRbfox1-iso1#1 or -iso#2. Analyses were done as in (a).

Back to article page