Figure S1. Validation of YKL-40 staining procedure. Macrophages were gated based on forward scatter (FS) and side scatter (SS) log parameters as shown in the left graph. The arrow points to analysis of macrophages that were stained using different protocols. The histogram shows YKL-40 staining (black-dashed line) of macrophages, which is easily distinguishable from the following control stains: YKL-40 is not detected without intracellular staining (ICS) permeablization of the cell (dark gray solid line). Unstained, permeabilized macrophages are similar to the other controls (filled dark gray peak). Staining of permeabilized macrophages with an irrelevant biotin-conjugated antibody (filled black peak) (A). Histogram shows the distinction of macrophages that were: not treated with GolgiStop and stained with YKL-40 (filled light gray peak), stained with anti-biotin antibody only (filled dark gray peak), stained with 0.5 µg anti-YKL-40 biotin-conjugated antibody (dashed line) and stained with 5 µg anti-YKL-40 biotin-conjugated antibody (black line) (B). Expression of YKL-40 was not observed in CD3+ lymphocytes (C).

Figure S2. Validation of TNFα secretion in cocultured macrophages. Astrocytes were plated in chamber slides and were grown for 2 days before macrophages were added to the culture. Cocultured cells were grown for 14 days and the culture medium was collected every 2 days and analyzed for TNFα. Time course of TNFα levels released into the culture medium was determined by TNFα ELISA kit. Data shown are from a single experiment using duplicate samples and are representative of three independent experiments.

BPA_550_sm_FigS1.tif684KSupporting info item
BPA_550_sm_FigS2.tif335KSupporting info item

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