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. 2022 May 10;61:101514. doi: 10.1016/j.molmet.2022.101514

Figure 5.

Figure 5

Increased cholesterol accumulation in Apoe−/−/HMGCR KI HSPCs contributes to HSPC mobilization and Ly6-Chi monocytosis. (A-I) Apoe−/− and Apoe−/−/Hmgcr KI mice were aged to 20 weeks old on a standard chow diet. (A) BM HSPCs were sorted using FACS before ex vivo cholesterol synthesis was measured via 14C-acetate labelling and TLC fractionation. n = 4 independent experiments. BODIPY-Cholesterol levels were measured in (B) BM HSPCs and (C) blood HSPCs. BM HSPC (D) proliferation and (E) CBS were measured via flow cytometry. BODIPY-Cholesterol levels were measured in (F) splenic HSPCs, and (G) splenic HSPC proliferation and (H) CBS were measured via flow cytometry. BODIPY-Cholesterol levels were measured in (I) blood Ly6-Chi monocytes. (J) Experimental overview. (K-L) Representative flow plots, and ratio of CD45.2 to CD45.1 cells in (K) blood Ly6-Chi monocytes and (L) BM HSPCs. (M) Graphic abstract. Results are means ± SEM and analysed using a Student unpaired t test, n = 4–9 per group (∗P < 0.05, ∗∗P < 0.01 when comparing Apoe−/− versus Apoe−/−/Hmgcr KI or Wt/Wt versus Hmgcr KI/Wt).