Figure 6.
Figure 6. Microvascular permeability is increased in mice with DSS-induced colitis with HYAL2-deficient platelets. (A) Distal colon sections from mice treated with 2.5% DSS for 7 days with or without transfusion of platelets were evaluated for HA-HC by immunostaining for HA (green) and the HCs of IαI (red) and counterstained with 4′,6-diamidino-2-phenylindole for nuclei. L denotes the lumen. Lower panels show the submucosal regions at higher magnification. Images are representative of at least 8 mice. (B) Measurement of HA levels in colon tissue from control, DSS-treated mice, and DSS-treated mice receiving platelets. (C) Quantification of colocalization of HA with HCs of IαI as a measurement of HA-HC deposition. (D) Measurement of EB extracted from distal colon tissue of mice treated with or without 2.5% DSS. Data are reported as mean ± SEM; n = 10 mice per group. *P < .05; **P < .01; ***P < .001. Imaging, detection, and software details: TCS SP5 II confocal/multiphoton high-speed upright microscope, HCX PL APO 40X/1.25NA oil immersion objective, HyD system detector, and LAS AF software (all Leica Biosystems). Pearson’s correlation coefficients were obtained by analyzing individual images (layers) of the Z-stack with Image-Pro Plus software.

Microvascular permeability is increased in mice with DSS-induced colitis with HYAL2-deficient platelets. (A) Distal colon sections from mice treated with 2.5% DSS for 7 days with or without transfusion of platelets were evaluated for HA-HC by immunostaining for HA (green) and the HCs of IαI (red) and counterstained with 4′,6-diamidino-2-phenylindole for nuclei. L denotes the lumen. Lower panels show the submucosal regions at higher magnification. Images are representative of at least 8 mice. (B) Measurement of HA levels in colon tissue from control, DSS-treated mice, and DSS-treated mice receiving platelets. (C) Quantification of colocalization of HA with HCs of IαI as a measurement of HA-HC deposition. (D) Measurement of EB extracted from distal colon tissue of mice treated with or without 2.5% DSS. Data are reported as mean ± SEM; n = 10 mice per group. *P < .05; **P < .01; ***P < .001. Imaging, detection, and software details: TCS SP5 II confocal/multiphoton high-speed upright microscope, HCX PL APO 40X/1.25NA oil immersion objective, HyD system detector, and LAS AF software (all Leica Biosystems). Pearson’s correlation coefficients were obtained by analyzing individual images (layers) of the Z-stack with Image-Pro Plus software.

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