Figure 2.

Effect of radiation and pUC on migration and cytoskeleton of non-GICs and GICs. A, U251 and 5310 non-GICs and GICs were irradiated with 5 and 10 Gy at 24 and 48 h. Cell lysates were extracted and analyzed by SDS-PAGE to determine the expression levels of uPAR and cathepsin B with and without radiation. GAPDH served as a loading control. All the results are representative of three individual experiments. B, U251 and 5310 non-GICs and GICs were transfected with pSV and pUC with and without irradiation for 24 and 48 h. Cell lysates were prepared and analyzed for uPAR and cathepsin B expression levels using western blotting. C, The protein expression levels of uPAR and cathepsin B in U251 and 5310 cells treated with pUC and radiation alone and in combination were analyzed by densitometric analysis and are depicted in the graph as relative protein expression (control of each set as 100%). D, Wound healing assay was performed with pSV and pUC and/or radiation treatments in U251 and 5310 non-GICs and GICs as described in Materials and methods. The cells were then fixed, stained with DAPI, and imaged using a fluorescence microscope (Olympus IX71, USA). Scale bar, 500 μm. E, The migration and wound healing capacities of U251 and 5310 non-GICs and GICs treated under the previously described conditions were measured using a microscope, analyzed, and are graphically represented as a relative percentage of wound healing (migration of pSV-treated samples as 100%). F, U251 and 5310 non-GICs and GICs treated with pSV and pUC with and without radiation were grown on fibronectin-coated 4-well chamber slides. The cells were then fixed with 4% buffered formalin, stained with vinculin antibody, and incubated with DAPI for a brief period of time before mounting. Scale bar, 200 μm. Values are mean ± SD of three different experiments (^*p<0.05, ^**p<0.01, in comparison with the control).