Figure 5

Bi-phasic response of SDIM1 in NT2 and mouse primary neurons treated with cell death-inducing stress. A. Changes in mRNA levels of SDIM1 in NT2 neurons treated with OGD and OGD with 16 h recovery were determined by qRT-PCR. The samples were measured against the cDNA of untreated NT2 neurons, set at 100%. Percentage of each treated sample was calculated by 100 × 2^-ΔCt, where ΔCt is the cycle number difference between treated sample and the control sample. The experiments were performed in triplicate. Asterisks indicate a significant difference (p-value = 6.25E-05 for control and 2hOGD; p-value = 0.0032 for control and 2hOGD + 16hR; p-value = 0.0023 for 2hOGD and 2hOGD + 16hR). B. a. Changes in SDIM1 protein levels in NT2 neurons treated with OGD and OGD plus 16 h recovery were determined by Western blotting with anti-SDIM1 antibody. Band intensity was measured using ImageJ and the level of SDIM1 was normalized against the level of β-actin and plotted in b. Asterisk indicates a significant difference (ρ < 0.005; t-test). C. a. Changes in SDIM1 protein levels in mouse primary neurons treated with OGD and OGD plus 16 h recovery were determined by Western blotting with anti-SDIM1 antibody. Band intensity was measured using ImageJ and the level of SDIM1 was normalized against the level of β-actin and plotted in b. Asterisk indicates a significant difference (ρ < 0.005; t-test). D. Changes of immunoreactivity of SDIM1 in primary neurons treated with OGD and OGD plus 16 h recovery. Cells were stained with anti-SDIM1 antibody. Cy3-conjugated anti-rabbit IgG was used to detect the specific immunostaining. The stained cells were viewed with a Zeiss Axiovert 200 M fluorescence microscope. Scale bar = 50 μM.