The process of Western Blotting is prone to error. Gels may not run correctly as a result of improper loading, transfers may not function correctly as a result of improper placement of components such as the transfer membrane and filter paper, and improper buffers may otherwise impede results. If the chemiluminescence kit does not function correctly as a result of overexposure or underexposure, an image will not be discernible. As a result of such factors, little besides the ladder and noise were noticeable on membranes after imaging. Results were obtained from data procured by the mentoring Ph.D. student, Mr. Zoltan Torok.
However, of those results displayed, no statistically significant differences were observed. This may be due to several factors, including that creatine monohydrate supplementation was not concentrated enough to produce any significant effects. The time between doxorubicin treatment and sacrifice may indeed be another factor. Several trends, however, are visible.
In the soleus, an increasing trend is seen in MyoD and Mrf4 expression while a decrease in Myf5 and myogenin expression was noticed with DOX treatment. The trend of CR+DOX, within the soleus, increased Myf5 and myogenin expression when compared to both CON and DOX. It is of interest that MyoD optical density was greater than both control and creatine & doxorubicin densities in the doxorubicin-treated group. Furthermore, it is of interest that Myf5 optical density exceeds both the resulting Myf5 optical densities of the creatine & doxorubicin as well as doxorubicin treatment groups in the EDL.
Further studies will likely involve Sprague-Dawley rats fed with greater concentrations of creatine as well as a shorter/longer? timespan from doxorubicin/saline injection to sacrifice.