Figure 3
Time-dependent changes in C/EBP mRNA expression following nerve injury. Real-time PCR was performed with specific primers for C/EBP between sham-operated and crush-operated L. stagnalis CNS at 1 hr, 3 hrs, and 5 hrs post injury. (A) Relative mRNA levels of C/EBP vs GADPH increased significantly following CNS injury as compared to the sham-operated controls at 3 hrs (2.35 ± 0.26, n = 5) (t = -3.5, df = 8, pP < 0.05). (B) Standard curves of C/EBP and GAPDH from 6 independent samples including sham-operated and crush-operated L. stagnalis CNS at 1 hr, 3 hrs, and 5 hrs post injury. The PCR efficiency of the primers was estimated by the slope (m) = -1/log(efficiency). The expression GAPDH was unchanged following CNS injury, and therefore CAPDH was used as an internal control. (C) Representative Ct-Ct correlation plots between C/EBP and the control gene, GADPH, at three different time points following injury. C1: 1 hr; C2: 3 hrs; C3: 5 hrs. Relative expression ratio between C/EBP and GADPH was estimated as Yintercept = -log(ratio)/log(efficiency C/EBP). (D) Relative gene expression of C/EBP vs. GADPH normalized to corresponding control samples. C/EBP increased in a time-dependent manner. 1 hr: 7.36 ± 0.73 (n = 5); 3 hrs: 2.35 ± 0.26 (n = 5), and 5 hrs: 1.98 ± 0.75 (n = 5). * indicates significant differences: ANOVA: F(2,12) = 17.4, p < 0.05. All data are presented as mean ± S.E.M.