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In the original article, the keys for the histograms in Fig. 3B and 3E were incorrect. The representative colors for cffDNA CM and cffDNA CM +TNF-α Block in panel (B) and cffDNA CM and cffDNA CM + IL-1β Block in panel (E)” were reversed. Fig. 3 is shown below with the corrected keys for panels (B) and (E). The figure legend was correct as published and is shown below for reference. Fig. 3 has been corrected in the online version of the article, which now differs from the print version as originally published.
FIGURE 3.
The cytokine response to cffDNA can induce endothelial activation. (A) Third trimester maternal PBMCs were stimulated overnight with Effectene alone or Effectene with cffDNA. CM from stimulated PBMCs (either CM or cffDNA CM) were harvested for overnight incubation with HUVECs. For designated wells, neutralizing Ab blocking TNF-α was added to supernatants from PBMCs treated with cffDNA CM. (B) Histograms from ICAM-1 and E-selectin staining of HUVECs from one representative experiment in which 10,000 single cells were collected for analysis. (C) Representative images of stained HUVECs with each treatment using ImageStream. (D) Third trimester maternal monocytes stimulated overnight with Effectene alone or Effectene with cffDNA. Supernatants from stimulated monocytes (either CM or cffDNA CM) were harvested for overnight incubation with HUVECs. For designated wells, neutralizing Ab blocking IL-1β was added to supernatants from monocytes treated with cffDNA CM. (E) Histograms from ICAM-1 and E-selectin staining of HUVECs from one representative experiment in which 10,000 single cells were collected for analysis. (F) Representative cell images of stained HUVECs using ImageStream. For all experiments, 10,000 single cells were collected and analyzed per replicate from each experiment (n = 2–3 donors). (A) and (D) represent median and IQR of the data. The p values were compared by Friedman test, with correction using false discovery rate for multiple comparison testing and two-stage linear step-up procedure of Benjamini, Krieger, and Yekutieli. Significant comparisons (p ≤ 0.05) are indicated by CM versus cffDNA CM (≠), cffDNA CM versus TNF-α or IL-1β block (Δ), and CM versus TNF-α or IL-1β block (φ).