Journal: PLoS ONE

Article Title: The Alarmin Concept Applied to Human Renal Transplantation: Evidence for a Differential Implication of HMGB1 and IL-33

doi: 10.1371/journal.pone.0088742

Figure Lengend Snippet: Confluent (≈95%) monolayer HUVEC were exposed to sixteen hours hypothermia/hypoxia in UW solution (H16h) followed by 1 hour (R1h), or 3 hours (R3h) of re-oxygenation in a new culture medium (Medium 200) at 37°C in 20% O 2 . Confluent (≈95%) monolayer HUVEC were used as controls (Ctl). Early release of HMGB1 and IL-33 by HUVEC in response to in vitro hypoxia/re-oxygenation (A–B). HMGB1 (A) and IL-33 (B) in cell culture supernatants were quantified by ELISA. Increase of IL-33 but not HMGB1 mRNAs in HUVEC in response to in vitro hypoxia/re-oxygenation (C–D). Total RNA was extracted from monolayer HUVEC at the indicated time points and expression of HMGB1 (C) and IL-33 (D) mRNAs was quantified by RT-qPCR. Data are expressed as means ± SEM or of fold change relative to D0 and are representative of three separate experiments. *p<0.05, **p<0.01, ***p<0.001 vs Ctl by Mann-Whitney test.

Article Snippet: Normal primary HUVEC up until passage 4 were cultured in Medium 200+ Low Serum Growth Supplement containing 2% fetal bovine serum (FBS) (Invitrogen, Saint Aubin, France) in a humidified atmosphere at 5% CO 2 and 37°C.

Techniques: In Vitro, Cell Culture, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, MANN-WHITNEY