Journal: bioRxiv
Article Title: Interleukin-6 Regulates the Neutrophil Response to Diverse Bacteria
doi: 10.64898/2026.01.08.698217
Figure Lengend Snippet: Representative maximum intensity projections of confocal images of HUVECs seeded in microfluidic devices and incubated with 0, 10, and 100 ng/mL IL-6 for 2 hours (A) with no bacteria and (C) with P. aeruginosa . Cells were fixed and stained with Hoechst (nuclei, blue), phalloidin (actin, red), and anti-ICAM-1 (neutrophil tight binding to endothelial cells, green) (scale bar = 250 μm). Images were thresholded in ImageJ to visualize the differences in ICAM-1. Representative raw images used for protein expression analysis can be found in Figure S2. (B, D) Fold change of ICAM-1 expression was calculated by dividing the total fluorescence intensity of un-thresholded images for each condition by the total fluorescence intensity for the 0 ng/mL IL-6 condition of the corresponding biological replicate. Fold change values for each biological replicate were averaged together and then all IL-6 conditions were compared to each other at each time point. Conditions at each time point were compared using a one-way ANOVA and then p-values were determined using Tukey’s HSD post-hoc test. Error bars indicate the mean ± SEM. (B) Data quantified from 9 devices for 0 ng/mL, 8 devices for 10 ng/mL, and 7 devices for 100 ng/mL across 4 independent experiments for protein expression when not exposed to bacteria. (D) Data quantified from 8 devices for 0 ng/mL, 9 devices for 10 ng/mL, and 8 devices for 100 ng/mL across 3 independent experiments for protein expression when exposed to P. aeruginosa .
Article Snippet: After 2 days of culture, 0, 10, 100 ng/mL IL-6 (130-093-929, Miltenyi Biotec) is added to the lumens and incubated at 37 °C and 5.0% CO 2 for approximately 45 minutes.
Techniques: Incubation, Bacteria, Staining, Binding Assay, Expressing, Fluorescence