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. Author manuscript; available in PMC: 2016 May 1.
Published in final edited form as: Nat Med. 2015 Oct 12;21(11):1307–1317. doi: 10.1038/nm.3960

Figure 5. Thrombin-PAR1 signaling induces NO production and HSC mobilization.

Figure 5

(a) NO levels in SLAM cells obtained from the bone marrow and peripheral blood. NOlow gating represents baseline reactivity; n = 3. (b) FACS analysis of eNOS Ser1177 phosphorylation in BM LSK cells 30 minutes following thrombin injection; n = 4. (c) Kinetics of eNOS phosphorylation on Ser1177 and Thr495 in bone marrow LSK cells following in vitro thrombin stimulation; n = 4. (d) Circulating LSK cells and bone marrow EPCR+ LSK cells following thrombin injection in mice pretreated with or without l-NAME; n = 6, P values, one-way ANOVA with Tukey’s post-test. (e) Circulating LSK cells and bone marrow EPCR+ LSK cells in wild-type or Nos3−/− bone marrow chimeric mice following thrombin injection; n = 4. (f) Circulating LSK cells and bone marrow EPCR+ LSK cells 60 minutes after SNAP injection; n = 4. (g) Cdc42 (purple) distribution on bone marrow EPCR+ (yellow) LSK cells 10 minutes following in vivo thrombin treatment. Scale bar, 7 µm. (h) FACS analysis of Cdc42-GTP following in vitro thrombin treatment for 15 minutes of bone marrow SK SLAM cells that had been pretreated with or without l-NAME for 2 hours; n = 4, P values, one-way ANOVA with Tukey’s post-test. (i) Cdc42-GTP in bone marrow SK SLAM cells (left) and LDV-FITC binding to bone marrow EPCR+ SK SLAM cells (right) following in vitro treatment with SNAP for 15 minutes; n = 4.