Disruption of NAD+ binding site in glyceraldehyde 3-phosphate dehydrogenase affects its intranuclear interactions

Figure 5 Nuclear accumulation of wild type and mutated enhanced green fluorescent protein-glyceraldehyde 3-phosphate dehydrogenase in live HCT116-4016 cells after transient transfections with plasmids coding for enhanced green fluorescent protein fusions with WT-glyceraldehyde 3-phosphate dehydrogenase, T99I-glyceraldehyde 3-phosphate dehydrogenase, and T99A-glyceraldehyde 3-phosphate dehydrogenase polypeptides. A: Distribution of EGFP-GAPDH variants in HCT116-4016 cells before (upper images) and after (lower images) treatment with 1 µmol/L araC for 24 h; B: Quantitative analysis of images shown in A was performed using ImageJ 1.48 v software (NIH, United States). For statistical evaluation, pixel analysis of 20-25 cells was performed for each experiment (mean ± SE). ¹Significant (P < 0.03) alteration in nuclear GFP-GAPDH between cells treated with araC and untreated cells. EGFP: Enhanced green fluorescent protein; GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; GFP: Green fluorescent protein.