Coadministration of HU and Akti XII efficiently inhibits neutrophil adhesion and platelet-neutrophil aggregation in venules, improves survival, and impairs neutrophil transmigration into alveoli in TNF-α-challenged Berkeley mice. TNF-α was intraperitoneally injected into Berkeley mice to induce severe inflammatory conditions. Intravital microscopy was performed as described in supplemental Methods. Neutrophils and platelets were labeled by infusion of Alexa Fluor 647-conjugated anti-Gr-1 and Dylight 488-conjugated anti-CD42c antibodies. (A) Timeline for the treatment and surgery (jugular cannulation and cremaster muscle exposure) in Berkeley mice. White arrows show direction of blood flow. (B) Representative images of intravital captures at various time points. Time 0 was set as the image capture was initiated at each vessel. (C-D) Number of rolling and adherent neutrophils. (E) The integrated median fluorescence intensities of anti-CD42c antibodies (F platelets) were normalized to the number of adherent neutrophils and plotted as a function of time. Data were obtained from 45-57 venules in 6-8 mice per group. (F) Survival curves of Berkeley mice during or after intravital microscopy. (G-H) Representative images from histochemistry of lung sections. The number of transmigrated neutrophils (arrow heads) was quantified in the field of view (110 mm²). Data represent the mean ± standard deviation (n = 25-30 sections in 6-8 mice per group). Bar = 20 μm. The survival rate was assessed with Mantel-Cox log-rank test. *P < .05, **P < .01, and ***P < .001 vs vehicle control; analysis of variance (ANOVA) and Dunnett’s test. #P < .05, ##P < .01, and ###P < .001 between two groups; Student t test.