Figure 2.
Genetic and pharmacologic inhibition of platelet PA alleviates venous thrombus formation. (A) Schematic of affected signaling pathways of platelet PA in PF4cre-TMEM16Ffl/fl mice. (B) Cell counts of peripheral blood for the indicated cell types and hematocrit quantification. Student t-test, 2-tailed, unpaired. (C) Experimental scheme of DVT in PF4cre-TMEM16Ffl/fl mice as well as thrombus weight (plotted) and incidence (percentage below graph) for n = 11 Cre– and n = 14 Cre+ PF4cre-TMEM16F mice. (D) Representative confocal image of IVC thrombi retrieved from Cre– or Cre+ mice. Scale bar = 100 μm. See supplemental Figure 5D-E for unstained examples of the same thrombi. (E) Quantification of PS-, CD41-, and Fbg-positive areas relative to total thrombus area. (F). Schematic of affected signaling pathways of platelet PA in PF4cre-CypDfl/fl mice. (G) Cell counts of peripheral blood for the indicated cell types and hematocrit quantification. (H) Experimental scheme of DVT in PF4cre-CypDfl/fl mice as well as thrombus weight (plotted) and incidence (percentage below graph) for n = 8 Cre– and n = 13 Cre+ PF4cre-CypD mice. (I) Schematic of affected signaling pathways of platelet PA in wild-type mice treated with the CA inhibitor methazolamide (MZA), which blocks aquaporin-1 (AQP-1)–mediated water influx into the ballooning platelet. (J) Flow cytometry–based measurement of percentage of procoagulant platelets and platelet MFIs for surface PS (measured through annexin V) in peripheral blood samples. Right: hematocrit measurements for mice from both experimental groups. (K) Experimental scheme of DVT and pharmacologic ablation of platelet PA using repetitive intraperitoneal injections of MZA in wild-type mice. (L) Thrombus weight in n = 14 vehicle- and n = 15 MZA-treated Bl6 mice. (M) Representative confocal image of IVC thrombi retrieved from vehicle- or MZA-treated mice. Scale bar = 500 μm. See supplemental Figure 6C for unstained controls. (N) Quantification of CD41- and Fbg-positive areas relative to total thrombus area. (O) Quantification of bleeding times of Bl6 mice undergoing tail section after pretreatment with vehicle or 20 mg/kg BW MZA. Unless otherwise stated, all statistical tests were Student t-test, 2-tailed, unpaired. P values corresponding to asterisks: ∗P < .05, ∗∗P < .01, ∗∗∗P < .005. For the schemes shown in A, F, and I, supplemental Figure 4A-B depicts a more detailed representation of signaling cascades implicated in platelet PA.

Genetic and pharmacologic inhibition of platelet PA alleviates venous thrombus formation. (A) Schematic of affected signaling pathways of platelet PA in PF4cre-TMEM16Ffl/fl mice. (B) Cell counts of peripheral blood for the indicated cell types and hematocrit quantification. Student t-test, 2-tailed, unpaired. (C) Experimental scheme of DVT in PF4cre-TMEM16Ffl/fl mice as well as thrombus weight (plotted) and incidence (percentage below graph) for n = 11 Cre– and n = 14 Cre+ PF4cre-TMEM16F mice. (D) Representative confocal image of IVC thrombi retrieved from Cre– or Cre+ mice. Scale bar = 100 μm. See supplemental Figure 5D-E for unstained examples of the same thrombi. (E) Quantification of PS-, CD41-, and Fbg-positive areas relative to total thrombus area. (F). Schematic of affected signaling pathways of platelet PA in PF4cre-CypDfl/fl mice. (G) Cell counts of peripheral blood for the indicated cell types and hematocrit quantification. (H) Experimental scheme of DVT in PF4cre-CypDfl/fl mice as well as thrombus weight (plotted) and incidence (percentage below graph) for n = 8 Cre– and n = 13 Cre+ PF4cre-CypD mice. (I) Schematic of affected signaling pathways of platelet PA in wild-type mice treated with the CA inhibitor methazolamide (MZA), which blocks aquaporin-1 (AQP-1)–mediated water influx into the ballooning platelet. (J) Flow cytometry–based measurement of percentage of procoagulant platelets and platelet MFIs for surface PS (measured through annexin V) in peripheral blood samples. Right: hematocrit measurements for mice from both experimental groups. (K) Experimental scheme of DVT and pharmacologic ablation of platelet PA using repetitive intraperitoneal injections of MZA in wild-type mice. (L) Thrombus weight in n = 14 vehicle- and n = 15 MZA-treated Bl6 mice. (M) Representative confocal image of IVC thrombi retrieved from vehicle- or MZA-treated mice. Scale bar = 500 μm. See supplemental Figure 6C for unstained controls. (N) Quantification of CD41- and Fbg-positive areas relative to total thrombus area. (O) Quantification of bleeding times of Bl6 mice undergoing tail section after pretreatment with vehicle or 20 mg/kg BW MZA. Unless otherwise stated, all statistical tests were Student t-test, 2-tailed, unpaired. P values corresponding to asterisks: ∗P < .05, ∗∗P < .01, ∗∗∗P < .005. For the schemes shown in A, F, and I, supplemental Figure 4A-B depicts a more detailed representation of signaling cascades implicated in platelet PA.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal