Fig. 5.

PGN treatment suppressed changes of cytokine levels caused by chronic stress. Six to eight-week-old wild type (C57BL/6) male mice were treated with PGN and stress as described in Fig. 1. After 12 h of stress, mouse spleens were harvested, and CD4⁺ T cells were isolated and cultured with anti-CD3/anti-CD28 antibodies in vitro for 48 h. RNA was extracted from CD4⁺ T cells and subsequently quantitative real-time PCR was performed. Cell-free supernatants were collected for subsequent ELISA assay.