Fig. 2.

Generation and characterization of HMGB1ECKO mice. A, Schematic diagram of the transgenic mice used to generate HMGB1ECKO mice. B, PCR analysis for HMGB1ECKO mice genotype. C, IF stain of HMGB1 (red) and CD31 (white) in aortas from HMGB1ECKO and control mice. Scale bar = 50 μm. D. Characterization of MLECs. Subconfluent MLECs (third passage) grown on glass cover slips were stained with endothelial surface markers, vWF, using anti-vWF antibody (primary) and a FITC-labeled secondary antibody. eNOS antibody (primary) and a Cys 3 secondary antibody. Scale bar = 20 μm. E. Representative Western Blot of HMGB1 expression in MLEC from control and HMGB1ECKO mice. Data are the average of triplicates from single experiments that were independently repeated 3 times. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)