Fig. 6.

VSL#3 CM stimulates TNF-α production and activates NF-κB in small intestinal organ cultures. Ilea from SAMP mice were harvested and cultured for 24 h with either VSL#3 CM, FE, or combination CM+FE and compared with controls (media alone). (A) Secreted TNF-α production was measured from resulting supernatants by ELISA and showed significantly increased levels in CM and CM+FE compared with controls. (B) Cytoplasmic and nuclear extracts analyzed by Western blot showed increased nuclear p65 (RelA) and decreased cytoplasmic IκBα levels in CM and CM+FE compared with controls, indicating activation of the NF-κB pathway by VSL#3. Data are representative of three independent experiments with similar results. *P < 0.001.