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. Author manuscript; available in PMC: 2016 Aug 15.
Published in final edited form as: JCI Insight. 2016 Aug 4;1(12):e87315. doi: 10.1172/jci.insight.87315

Figure 5. Intravital 2-photon imaging demonstrates impaired neutrophil extravasation into TLR9-deficient heart grafts.

Figure 5

Neutrophil tra_cking in (A) TLR2-deficient (see Supplemental Video 10; n = 4 mice), (B) toll-interleukin 1 receptor domain containing adaptor protein–deficient (TIRAP-deficient; see Supplemental Video 11; n = 4 mice), and (C) TLR9-deficient (see Supplemental Video 12; n = 4 mice) cardiac grafts. White arrows in A and B point to sites of neutrophil extravasation. Relative time is displayed in hrs:min:sec. Scale bars: 50 μm. (D) Percentage of neutrophils that entered myocardial tissue during imaging period was comparable between WT, TLR2-deficient, and TIRAP-deficient hearts. However, the percentage of extravasated neutrophils was significantly lower in TLR9-deficient than WT cardiac grafts. (E) Neutrophil rolling velocities were significantly higher in coronary veins of TLR9-deficient cardiac grafts when compared with WT hearts. Rolling velocities in TLR2- or TIRAP-deficient hearts were comparable with WT cardiac grafts. (F) Intraluminal crawling velocities of neutrophils did not di_er significantly between WT, TLR2-deficient, and TIRAP-deficient hearts but were significantly lower than WT conditions when hearts lacked expression of TLR9. *P < 0.05; **P < 0.01 (one-way ANOVA). Data in D, E, and F are derived from 4 mice for each experimental group. For D, E, and F, symbols represent averages obtained from individual mice with over 30 neutrophils examined per mouse, horizontal bars denote means, and error bars denote ±SEM.