Analysis of cell cycle specific genes on mRNA and protein level.

<p>The expression of p53 was analysed by microarray, qPCR and Western blot. (A) IPEC-J2 showed a 27.4 fold increase of the p53-mRNA level in comparison to IPEC-1. This was confirmed by qPCR and a 300-fold increase was detected in IPEC-J2. On the other hand, no protein expression was observed in IPEC-1/IPEC-J2 and Caco-2 as well as in the cells, which were treated with the p53-activator. In the next step, the expression of BAX, BAD and BCL-X was analysed using microarray, qPCR and Western blot. (B) BAD showed a 1.8 fold increase in IPEC-J2 in the microarray. Same results were found in qPCR, but no differences in the protein expression were found between the cell lines. A significant BCL-X up-regulation was detected in the microarray in IPEC-J2 but no differences were present between IPEC-1 and IPEC-J2 in the Western blot analyses. No BAX-protein was observed in both cell lines. When the total RNA and protein content was examined, significant differences were found between the cell lines. IPEC-J2 showed a higher RNA- and protein content in comparison to IPEC-1. RPL10A as important gene of the 60S ribosomal subunit was significantly decreased in the microarray but not in qPCR in IPEC-J2 in comparison to IPEC-1. Western blot analyses showed a slightly decrease of the protein in IPEC-J2 over time. (C) The RNA and protein content per cell was measured in both cell lines. IPEC-J2 showed a significant higher level in RNA and proteins. On the other hand, a slight decrease on protein level of RPL10A was found in IPEC-J2 in comparison to IPEC-1 in Western blot analyses.