Fig 1. Time course of LPS uptake by hepatocytes.

(A) For a live-imaging experiment fresh isolated primary hepatocytes were plated on 35 mm glass bottom dish precoated with collagen I (MatTek) and incubated overnight. The cells were stained with Hoechst to identify nuclei (blue) prior Alexa Fluor 488-conjugated E.coli LPS (green) addition and then immediately imaged constantly with the 63x objective using a Leica Inverted Fluorescence microscope, at 37°C and 5% CO2. (B) Time course of LPS uptake by HepG2 cells. HepG2 cells were treated with Alexa-488 FluorTM E.coli LPS (2.5 μg/ml) for 15, 30 min, 1, 2, 4 and 6 hours in media containing 20% human plasma. LPS uptake was measured by flow cytometry. Data presented as mean fluorescence intensity, mean±SEM (n = 3).