(a) NO (green) levels in BM LSK EPCR+ (purple)
and EPCR− lineage negative Sca-1+ (yellow)
c-Kit+ (red) cells. Scale bar, 7 µm. (b)
Percentage of NOlow SK SLAM cells following in vitro
treatment of bone marrow cells with EPCR neutralizing or non-neutralizing
antibody; n = 4. (c) Percentage of
NOlow cells and NO levels in wild-type or
Procrlow bone marrow LSK SLAM cells, with or
without in vitro aPC treatment for 1 hour; n =
3, P values, two-way ANOVA with Bonferroni post-test.
(d) Kinetics of eNOS phosphorylation on Ser1177 or Thr495 in
bone marrow SK SLAM cells following in vitro aPC stimulation;
n = 4. (e) eNOS phosphorylation on Thr495 in
bone marrow LSK SLAM cells following in vitro stimulation with
aPC for 15 minutes, active-site blocked aPC, FVIIa, or active-site blocked
FVIIa; n = 4, P values, one-way ANOVA with
Tukey’s post-test. (f) Circulating LSK cells in wild-type
or Procrlow chimeric mice following prolonged
l-NAME treatment for 5 days; n = 5, P values,
two-way ANOVA with Bonferroni post-test. (g) Adhesion and migration
of Procrlow bone marrow LSK cells following
in vitro
l-NAME or PBS treatment for 2 hours; n = 6.
(h) Circulating LSK (n = 8), bone marrow
EPCR+ SK SLAM cells (n = 4) and representative
FACS plot for EPCR expression following prolonged l-NAME or PBS
treatment for 5 days. (i) Competitive repopulation of bone marrow
cells obtained from donors treated with l-NAME or PBS, assessed 16
weeks following transplantation; each dot represents an individual recipient
mouse; P values, one-way ANOVA with Tukey’s post-test.
(j) Survival after 5-FU treatment, with or without SNAP
treatment (n = 9, *** P value = 0.0003), in
wild-type vs F2r−/− mice (n =
5, ** P value= 0.0010) or co-treated with EPCR neutralizing or
non-inhibitory antibody (n = 9, *** P value
< 0.0001).