Table 1.
Sequences of primers used for RT-PCR.
Figure 1.
Stable IGF-1-GFP expression in spinal cord-derived NSCs.
Phase-contrast image of pcDNA3.1-IGF-1/GFP-transfected NSCs selected by adding G418 for 14 days (A); fluorescence images of pcDNA3.1 (B) and pcDNA3.1-IGF-1-GFP transfected NSCs (C) at 24 h after transfection.
Figure 2.
Confirmation of IGF-1/GFP expression in NSCs.
RT-PCR analysis of IGF-1 in the pcDNA3.1 (lane 2) and pcDNA3.1-IGF-1/GFP-transfected NSCs (lane 3) (A). As shown in lane 1, DNA marker sizes were 100, 250, 500, 750, 1000 and 2000 bp from bottom to top. Similarly with the positive control (lane 4), the pcDNA3.1-IGF-1/GFP-transfected NSCs (lane 3) generate a 407 bp product that was identical with IGF-1 mRNA size. Western blot analysis of IGF-1 in the pcDNA3.1 (lane 1) and pcDNA3.1-IGF-1/GFP-transfected NSCs (lane 2) (B). IGF-1 protein can be detected in pcDNA3.1-IGF-1-GFP-transfected NSCs (lane 2) but not in the empty vector transfected cells (lane 1).
Figure 3.
Morphological appearance of differentiated NSCs.
Phase-contrast image of empty vector-transfected NSCs (A). Cells showed astrocyte-like morphology. Phase-contrast image of IGF-1-GFP expressing NSCs (B). Cells showed oligodendrocyte-like morphology.
Figure 4.
Expression of MPB in NSCs transfected with pcDNA3.1 (A) and pcDNA3.1-IGF-1/GFP (B). As shown, MBP expression increased dramatically in the IGF-1-GFP expressing NSCs.
Figure 5.
Effect of IGF-1 on the activation of ERK1/2.
Western blot analysis of pERK1/2 in NSCs at different time points (0, 0.5, 1 or 2 h) after transfection of pcDNA3.1-IGF-1/GFP (A and B), and in IGF-1/GFP expressing NSCs treated with different concentrations of PD98059 (0, 5, 10, 20 or 40 µM) (C and D). *p<0.05 denotes as statistically significant in comparison to all other groups.
Figure 6.
Effect of IGF-1 on the immediate genes of c-fos, c-jun and c-myc assessed by semi-quantitative RT-PCR analysis.
Expressions of c-fos, c-jun, c-myc in IGF-1-transfected NCSs (A and B); c-fos, c-jun expression in NSCs were determined in control, or after IGF-1 expression and ERK1/2 inhibitor treatment (B and D). *p<0.05 denotes as statistically significant in comparison to all other groups.