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. Author manuscript; available in PMC: 2014 Jul 21.
Published in final edited form as: Arterioscler Thromb Vasc Biol. 2013 Aug 29;33(11):2509–2517. doi: 10.1161/ATVBAHA.113.301989

Figure 2.

Figure 2

Treatment of whole aortas with proinflammatory factors before mouse aortic endothelial cell (MAEC) isolation effectively perturbs gene expression of inflammatory markers without altering endothelial cell (EC) marker levels. Relative transcription levels of EC marker genes and select stress–response genes in MAECs isolated after treatment with Dulbecco’s Modified Eagle’s Medium (DMEM) cell culture medium containing various additives, as determined by reverse transcription quantitative polymerase chain reaction and normalized to B2M. A, Expression levels of the EC markers von Willebrand factor (vWF), platelet/endothelial cell adhesion molecule 1 (CD31) in native MAEC preparations compared with MAECs isolated from aortas incubated in DMEM for 4 hours. No significant expression differences in EC cell marker expression after incubation in cell culture media were observed as determined by a t test. Error bars show SE from 4 biological replicates for each condition. B, EC markers and heme oxygenase 1 (Hmox1) in MAECs isolated from aortas incubated in DMEM compared with DMEM containing 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (oxPAPC) for 4 hours. Although the transcription levels of EC cell markers remained the same, there was a strong induction of Hmox1 in response to treatment with oxidized phospholipid. Error bars show SE from 3 biological replicates. t test: ***P <0.001. C, Expression levels of EC markers Hmox1 in MAECs isolated from aortas incubated in DMEM vs DMEM containing oxidized low-density lipoprotein (oxLDL) for 4 hours. We see an induction of Hmox1 but not EC markers in response to treatment. Error bars show SE from 3 biological replicates. t test: *P <0.05. D, Transcription levels of EC markers, Hmox1, and vascular cell adhesion molecule 1 (Vcam1) in MAECs isolated from aortas incubated in DMEM vs DMEM containing lipopolysaccharide (LPS) for 4 hours. No response was observed in Hmox1 expression, but Vcam1 transcript levels are induced as expected. Error bars show SE from 2 biological replicates. t test: *P <0.05.