Fig. 5.

Oxidative stress and estrogen deprivation promote higher production of IL-6 and IL-8 secretion in murine bone marrow stromal cells. ELISA assays demonstrated significantly higher levels of production of IL-6 (a and e) and IL-8 (b and f) in the conditioned medium from murine stromal layers treated with H₂O₂ and ICI 182780. Induction of hypoxia by CCCP did not cause similar effects (c and d). Near confluent stromal monolayers cultured in 24-well plates were treated with H₂O₂ and CCCP in serum-free medium for 1 h, washed once and incubated in DMEM/10%FCS medium. Supernatant samples were collected at 24, 48 and 72 h. Confluent stromal monolayers cultured in 24-well plates were treated with ICI 182780 in medium without phenol-red for 48 h and 72 h, washed once and incubated with DMEM/10%FCS medium. The IL-6 and IL-8 levels were determined using murine IL-6 and CXCL1/KC ELISA kit according to manufacturer’s instructions. Error bars: Standard Deviation. *p < 0.05, **p < 0.01 ***p < 0.001