Figure 5
Figure 5. Analysis of TFflox/flox, Tie-2Cre mice. (A) PCR analysis of mouse TF DNA isolated from blood cells demonstrates deletion of the TF gene. Arrows indicate PCR products for wild type and floxed alleles. A nonspecific band is present below the TF band. The mouse IL-2 gene was used as a loading control. WT indicates DNA from a WT mouse. (B) Real-time PCR analysis of TF mRNA in blood cells isolated from TFflox/flox and TFflox/flox,Tie-2Cre mice before (n = 2 per group) and after (n = 4 per group) LPS injection (2 hours). (C) PCR analysis of mouse TF DNA isolated from ECs demonstrates deletion of the TF gene. Arrows indicate PCR products for wild type and floxed alleles. The mouse IL-2 gene was used as a loading control. Presence or absence of Cre recombinase (Cre) in different mouse groups is shown. Low levels of fibroblast contamination likely explain the residual TF-positive band in 1 of the 4 cell preparations from TFflox/flox,Tie-2Cre mice[b]. (D) Plasma TAT levels were analyzed in TFflox/flox (n = 19; mean ± SEM 15.2 ± 1.7 ng/mL) and TFflox/flox,Tie-2Cre (n = 21; mean ± SEM 7.7 ± 0.9 ng/mL) mice 8 hours after LPS injection. *P < .001.

Analysis of TFflox/flox, Tie-2Cre mice. (A) PCR analysis of mouse TF DNA isolated from blood cells demonstrates deletion of the TF gene. Arrows indicate PCR products for wild type and floxed alleles. A nonspecific band is present below the TF band. The mouse IL-2 gene was used as a loading control. WT indicates DNA from a WT mouse. (B) Real-time PCR analysis of TF mRNA in blood cells isolated from TFflox/flox and TFflox/flox,Tie-2Cre mice before (n = 2 per group) and after (n = 4 per group) LPS injection (2 hours). (C) PCR analysis of mouse TF DNA isolated from ECs demonstrates deletion of the TF gene. Arrows indicate PCR products for wild type and floxed alleles. The mouse IL-2 gene was used as a loading control. Presence or absence of Cre recombinase (Cre) in different mouse groups is shown. Low levels of fibroblast contamination likely explain the residual TF-positive band in 1 of the 4 cell preparations from TFflox/flox,Tie-2Cre mice[b]. (D) Plasma TAT levels were analyzed in TFflox/flox (n = 19; mean ± SEM 15.2 ± 1.7 ng/mL) and TFflox/flox,Tie-2Cre (n = 21; mean ± SEM 7.7 ± 0.9 ng/mL) mice 8 hours after LPS injection. *P < .001.

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