Effect of EDCs alone and in combination on the secretion of TNF-α by differentiated THP-1 cells.

<p>Differentiated THP-1 cells were exposed to BPA, DEHP, DBP 4-OP or a combination of both for 24 hours. Culture medium was replaced by fresh medium containing 10 ng/ml of LPS for a subsequent 24 hours of incubation. TNF-α concentrations were determined using a sandwich ELISA test. Values are means ± SD (N = 3). Each experiment has been performed three times and in triplicate. a: <i>vs</i> control (Tukey’s test, p<0.05); b: <i>vs</i> combination (One-way ANOVA, Tukey post test, p<0.05).</p

Effect of EDCs alone and in combination on estrogen receptor protein expression and signaling.

<p>(A) Differentiated THP-1 cells (PMA 5 ng/ml for 48 hours) were exposed to ICI-182780 (1 μM), BPA, DEHP, BPA + DEHP, DBP, BPA + DBP, 4-OP or 4-OP + DEHP (each EDC at 0.1 μM) for 24 hours. Culture medium was replaced by fresh medium containing 10 ng/mL of LPS for a subsequent 24 hours of incubation. Whole cell lysates (20 μg) were separated by SDS-PAGE (10%) and immunoblotted with the anti-estrogen receptor antibody as described in Materials and Methods. The same blot was stripped and immunoblotted with control antibody (anti-β-actin). Histograms represent the relative quantifications of ERα which were performed in comparison with β-actin. Values in non-treated cells are taken as 100%. Each experiment has been performed four times. Values are means ±SD (N = 4). a: <i>vs</i> control (One-way ANOVA, Tukey post test, p<0.05). (B) Differentiated THP-1 cells (PMA 5 ng/ml for 48 hours) were exposed to ICI-182780 alone and in combination with BPA, DEHP, BPA + DEHP, DBP, BPA + DBP, 4-OP, 4-OP + DEHP for 24 hours. Culture medium was replaced by fresh medium containing 10 ng/mL of LPS for a subsequent 24 hours of incubation. Each EDC was also tested without ICI. EDCs and ICI were used at 0.1 μM and 1 μM respectively throughout this experiment. Whole cell lysates (20 μg) were separated by SDS-PAGE (10%) and immunoblotted with the anti-phospho-ERK antibody as described in Materials and Methods. The same blot was stripped and immunoblotted with anti-ERK antibody. Histograms represent the relative quantifications of phospho-ERK which were performed in comparison with ERK. Values in non-treated cells are taken as 100%. Each experiment has been performed three times. Values are means ±SD (N = 3). a: <i>vs</i> control (One-way ANOVA, Tukey post test, p<0.05); b: <i>vs</i> exposure to corresponding EDCs after treatment by ICI (One-way ANOVA, Tukey post test, p<0.05).</p

Effect of EDCs alone and in combination on phagocytosis by differentiated THP-1 cells.

<p>Differentiated THP-1 cells (PMA 5 ng/ml for 48 hours) were exposed to different concentrations (0.001, 0.1, 1 and 10 μM) of BPA, DEHP, DBP, 4-OP or a combination of both for 24 hours. Culture medium was replaced by fresh medium containing 10 ng/ml of LPS for a subsequent 24 hours of incubation. Panels A and B depict confocal laser microscopy analysis of phagocytosis of FITC-latex beads by untreated differentiated THP-1 cells (A) or by differentiated THP-1 cells exposed to BPA (B). Arrows show examples of phagocytized FITC-beads. Panel C represents the effects of BPA, DEHP, DBP, 4-OP and a combination of both (BPA + DEHP, BPA + DBP, 4-OP + DEHP) on phagocytosis of latex beads by differentiated THP-1 cells. Values are means ± SD (N = 3). Each experiment has been performed three times and in triplicate. a: <i>vs</i> control (One-way ANOVA, Tukey post test, p<0.05).</p

Effect of EDCs alone and in combination on the secretion of IL-1β by differentiated THP-1 cells.

<p>Differentiated THP-1 cells were exposed to different concentrations of BPA or DEHP or DBP or 4-OP or a combination of both for 24 hours. Culture medium was replaced by fresh medium containing 10 ng/mL of LPS for a subsequent 24 hours of incubation. IL-1β concentrations were determined using a sandwich ELISA test. Values are means ± SD (N = 3). Each experiment has been performed three times and in triplicate. a: <i>vs</i> control (Tukey’s test, p<0.05); b: <i>vs</i> combination (One-way ANOVA, Tukey post test, p<0.05).</p