Figure 4
Cell type specific expression and cellular localization of SDIM1. A. Changes in mRNA levels of SDIM1 during RA-induced differentiation of NT2 cells were determined by qRT-PCR. The samples were measured against the cDNA of undifferentiated NT2 cells as a control, set at 100%. Percentage of each sample was calculated by 100 × p 2-ΔCt, where ΔCt is the cycle number difference between test sample and the control sample. undiff - undifferentiated NT2 cells (control), NT2A - NT2 astrocytes, NT2N - NT2 neurons. The experiments were performed in triplicate. Asterisks indicate a significant difference (p-value = 0.00024 for Undiff and NT2A; p-value = 0.0027 for Undiff and NT2N; p-value = 0.0029 for NT2A and NT2N). B. Changes in SDIM1 protein levels were determined by Western blotting with anti-SDIM1 antibody using 100 μg/lane of total cellular protein. The Western blotting of β-actin was shown as loading control. C. Subcellular localization of SDIM1. Mouse primary cortical neurons were fixed and stained with anti-SDIM1 and anti-MAP2 antibodies. Staining with secondary anti-body alone was also performed as a negative control (image not show). FITC-conjugated anti-rabbit IgG (for SDIM1) and rhodomine-conjugated anti-mouse IgG (for MAP2) were used to detect the specific immunostaining. The nuclei were stained with DAPI and viewed with a Zeiss Axiovert 200 M fluorescence microscope. Arrowheads indicate glial cells or inmature neurons. Scale bar in c = 50 μM.