Fig. 4
LPS, But Not Cocaine, Induces Cytokine Production at the BBB Which Does Not Disrupt BBB Integrity. BBB models were treated with cocaine (10 μM, burgundy), LPS (100 ng/mL, fuchsia), or vehicle (teal) for 24 h, after which time (A-K) supernatants were collected for cytokine determination or (L) permeability to 4 kDa FITC-Dextran was performed. (A-K) Multiplexed cytokine analysis was performed by flow cytometry to evaluate (A) IL-1β, (B) IL-6, (C) TNF-α, (D) IP-10, (E) IFN-λ1, (F) IL-8, (G) IL-12p70, (H) IFN-α2, (I) IFN-β, (J) GM-CSF, and (K) IL-10. Dashed line denotes quantitative limit of detection for IL-8. (L) Four kDa FITC-Dextran was added to the apical portion of the BBB for 30 min at 37 °C, 5% CO2, the media in the bottom collected, and evaluated by a fluorescence microplate reader. FITC-Dextran alone (green) was used as a positive control to represent maximal BBB permeability. Six individual experiments were performed (represented by individual dots). Data are represented as mean ± standard deviation. **p < 0.01. ***p < 0.001. ****p < 0.0001. One-way ANOVA was performed