Figure 7.

Effect of the dual altered peptide ligand (APL) on the expression of apoptotic markers in lymph node (LN) cells of C57BL/6 mice immunized with Torpedo acetylcholine receptor (TAChR). C57BL/6 mice were either immunized with TAChR [10 µg per mouse in complete Freund's adjuvant (CFA)] and concomitantly administered subcutaneously (s.c.) with the dual APL/reversed dual APL [200 µg per mouse in phosphate-buffered saline (PBS)] or immunized with TAChR alone. Ten days after immunization, popliteal LN cells were harvested. (a) Total RNA was extracted from LN cells and real time polymerase chain reaction (PCR) was performed for caspase 8, caspase 3, cFLIP and Bcl-2 as described in Materials and methods. Results are expressed as the mean percentage of the gene expression of triplicates ± SD values. *P < 0·05, **P < 0·001 when gene expression in the presence of the dual APL was compared with that in the absence of the dual APL. (b) Lysates were prepared from LN cells, separated on sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a nitrocellulose membrane that was later blotted with anti-p-c-Jun-NH2-terminal kinase (JNK) or anti-p-38 or their respective totals. Results are expressed as densitometry units (measured with National Institutes of Health image software). Shown is one of two experiments performed.