After many repeates in real time experiment, I am getting a differential expression of Caspase 3, 9 and P53 genes (no dose response).
Relative to the solvent control a significant 164, 72.4 and 117 % differential up-regulation in the mRNA expression of p53 gene has been observed (Please see attachment Fig). For the caspase 3 and 9 genes, Up-regulation of mRNA expression has been noticed only at two initial doses (0.046, 0.092 mg /kg bw/day). However, the mRNA expression at the highest dose of 0.184 mg /kg bw/day has exhibited a decline in mRNA expression. Relative to control the expression of mRNA for caspase 3 and 9 were 120.7, 202.4, 67.5 and 186, 213, 123.2 % mRNA expression has been observed at 0.046, 0.092 and 0.184 mg /kg bw/day.
In order to confirm the translational product, I have also checked the enzyme activity of Caspase 3 and 9 (no p53). In this experiment 1.2, 1.6-fold and 2.7, 3.6-fold (p<0.05) increase in caspase 3 and 9 activity at 0.092 and 0.184 mg/kg bw/day treated animals has been recoreded.
As a general rule, if I would get an increase in mRNA expression of Caspase 3 and 9, with low enzyme or protein activity, this suggests a post-translational modification.
However, in my case this is reverse, I am getting the low mRNA expression at the highest dose but the enzyme activity at the same dose is higher.
Therefore, I am stucked at this point and need expert views from the leading workers to resolve my problem (if possible, with a relvant literature to support) which could lead me to draw a meaningful conclusion.
I am attaching the figure of mRNA expression and enzyme activity data for you.
Looking to hearing from you soon.....