Extended Data Fig. 9: Splenic endothelial cells have greater lipid availability and regulate HSPC activation.
From: Hyperlipidemia-induced hematopoiesis is repressed by MLKL in endothelial cells of the splenic niche
a, Representative flow cytometry plots and quantification of bone marrow and splenic lipid content (BODIPY+) from Apoe-/- mice after 4 weeks of HCD feeding and Mlkl knockdown. n = 4 mice per group. b, DiI-labelled acetylated low-density lipoprotein (DiI-acLDL) incorporation in bone marrow and splenic endothelial cells was assessed 4 hours after in vivo administration to 5-week HCD-fed Apoe-/- mice. n = 5 mice per group. c,d, Representative images and quantification of primary mouse splenic (c; siControl n = 6 replicates, siMLKL n = 5 replicates) and bone marrow (d; siControl n = 10 replicates, siMLKL n = 9 replicates) endothelial cells transfected with non-targeting (siControl) or Mlkl-targeting (siMLKL) siRNA. Data representative from 3 independent experiments. MLKL shown in purple and nuclei shown in blue. Scalebar = 10μm. Cells are outlined by the white dashed lines. e, Representative flow cytometry plots and quantification of pStat5 expression in CMFDA+ HSPCs after co-culture with transfected splenic endothelial cells. siControl n = 3 replicates, siMLKL n = 4 replicates. Representative of 3 individual experiments. f, Myeloid colony formation from HSPCs after co-culture in transwells with transfected splenic endothelial cells. n = 3 replicates per group. Data representative from 3 independent experiments. Data are shown as mean ± s.e.m. Statistical significance was determined by two-way ANOVA followed by Holm-Sidak’s post-hoc test for multiple comparisons (a), two-tailed paired t-test (b), two-tailed Mann Whitney test (c) or two-tailed unpaired t-test (d-f).
