Table 1.
Arterial blood gas analysis Pre- and Post-AMRT.
Figure 1.
Change in phosphoAktSer473 before (Pre) and after (Post) AMRT determined by immunohistochemistry.
(A) Bars show statistical analysis of gray values (au) between Pre and Post condition. The phosphoAktSer473 signal in human RBCs significantly increased from Pre to Post (P<0.05). (B) Pictures show representative phosphoAktSer473 staining before and after the test, respectively. Magnification for all images was 400-fold. Data in (A) are presented as mean ± S.E.M of n = 6.
Figure 2.
Change in detectable RBC-NOS and RBC-NOS phosphorylation before (Pre) and after (Post) AMRT determined by immunohistochemistry.
(A) Bars show statistical analysis of gray values (au) revealing no statistical difference in detectable total RBC-NOS between Pre and Post conditions. (B) shows representative detectable RBC-NOS staining before and after the test, respectively. (C) The data reveal that RBC-NOS phosphorylation at Ser1177 residue significantly increased from the Pre to Post situation (P<0.01). (D) Pictures show representative RBC-NOSSer1177 staining before and after the test, respectively. (E) The graph shows no statistical difference in gray values (au) between Pre and Post conditions for RBC-NOSThr495. (F) Pictures show representative RBC-NOSThr495 staining before and after the test, respectively. Magnification for all images was 400-fold. Data (A), (C) and (E) are presented as mean ± S.E.M of n = 6.
Figure 3.
Change in cGMP before (Pre) and after (Post) AMRT determined by immunohistochemistry.
(A) Bars show statistical analysis of gray values (au) between Pre and Post conditions. The cGMP signal in human RBCs significantly increased at the end of the test (Post) (P<0.05). (B) Pictures show representative cGMP stainings before and after the test, respectively. Magnification for all images was 400-fold. Data in (A) are presented as mean ± S.E.M of n = 6.
Figure 4.
Change in RBC-NOSSer1177 after PI3 kinase inhibition via wortmannin determined by immunohistochemistry.
(A) Bars show statistical analysis of gray values (au) before (0 min) and after a 30 min incubation of whole blood with PI3 kinase inhibitor wortmannin (c = 10 µM). RBC-NOSSer1177-signal significantly decreased after wortmannin incubation (P<0.01). (B) Pictures show representative RBC-NOSSer1177 staining before and after the test, respectively. Data in (A) is presented as mean ± S.E.M of n = 6.
Figure 5.
Change in phosphoAktSer473, RBC-NOS and cGMP before (Pre) and after (Post) AMRT determined by western blot analysis.
(A) Specific representative bands for phosphoAktSer473, total detectable RBC-NOS, RBC-NOSSer1177, RBC-NOSThr495 and cGMP migrated at 60kDa, 140kDa, 130kDa, 140kDa and 80kDa, respectively. PhosphoAktSer473, RBC-NOSSer1177 and cGMP obtained Post-AMRT were significantly increased compared to Pre-AMRT. Bands obtained from total detectable RBC-NOS and RBC-NOSThr495 remained unchanged. (B) Relative intensity obtained from western blot bands was significantly increased for phosphoAktSer473 (P<0.05), RBC-NOSSer1177 (P<0.001) and cGMP (P<0.05) Post-AMRT in relation to Pre- AMRT (set to “1”). Relative intensity of total detectable RBC-NOS and RBC-NOSThr495 remained unaltered. Data in (B) are presented as mean ± S.E.M of n = 6.
Figure 6.
NO-generation in human RBCs after exercise.
(A) DAF fluorometry revealed no detectable NO-generation Pre AMRT. (B) Strong NO generation signals were observed Post AMRT. (C) The NO generation was strongly reduced after AMRT by the PI3 kinase specific inhibitor wortmannin (c = 10 µM). Bar, 20 µm.
Figure 7.
Influence of AMRT on RBC deformability.
RBC deformability was obtained before (Pre) and after (Post) AMRT. (A) EImax significantly increased from Pre to Post exercise (P<0.05). (C) SS ½ significantly decreased Post AMRT. Data are presented as mean ± S.E.M of n = 15.
Figure 8.
Influence of wortmannin on RBC deformability.
RBC deformability was measured before and after 30 min incubation with wortmannin and PBS, as control. (A) No difference in EImax was observed after PBS treatment. After wortmannin incubation EImax was significantly decreased (P<0.05). (B) SS ½ was unchanged after PBS treatment but significantly increased after wortmannin application, indicating decreased deformability (P<0.05) Data are presented as mean ± S.E.M of n = 10.