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Figure 4. Reverse transcription-polymerase chain reaction (RT-PCR) and quantitative PCR (qPCR) analysis of select genes. Expression levels of a select set of genes found from the array screen were assayed by RT-PCR and qPCR to validate the array results. RT-PCRs were carried out for 25 cycles, except for those genes identified with an asterisk (30 cycles). Gene names or symbols are indicated to the left of each panel. Numbers below each panel represent the qPCR values for expression levels when normalized to ODC and with 1 day postamputation (1dPA) samples arbitrarily set to a value of 1. Tables to the right of each panel compare the ratios of gene expression determined by qPCR with those determined from the array screen. Columns A, B, and C refer to the designations outlined in Figure 1 and the number coloring is the same as described, e.g., green numbers for A indicate that the expression ratio is higher in stage 53 (st53) 1dPA compared to st57 1dPA. A dash indicates that there was no statistically significant ratio for that value from the array screen. ODC, ornithine decarboxylase; SOCS3, suppressor of cytokine signaling 3; FGL2, fibrinogen-like protein 2; C3, complement factor 3; nNOS, neuronal nitric oxide synthase.Download figure to PowerPoint |
