Figure 1. Loss of Bhlhe40 dysregulates the cell cycle in LPMs.

a, Flow cytometry of Bhlhe40^GFP transgene reporter expression in blood monocytes (representative of 2 experiments, n=5 Bhlhe40^GFP+, 2 Bhlhe40^GFP-); red pulp macrophages, microglia, Kupffer cells, kidney macrophages, SI macrophages, and peritoneal macrophages (representative of 2 experiments, n=4 Bhlhe40^GFP+, 2 Bhlhe40^GFP-); and AMs (representative of 3 experiments, n=6 Bhlhe40^GFP+, 3 Bhlhe40^GFP-) from Bhlhe40^GFP+ and Bhlhe40^GFP- mice. b, Flow cytometry of peritoneal macrophage subsets in Bhlhe40^+/+ and Bhlhe40^−/− mice (representative of 6 experiments, n=22/group). c, Numbers of LPMs as in b, SPMs (pooled from 5 experiments, n=19/group), AMs (pooled from 4 experiments, n=13 Bhlhe40^+/+, 12 Bhlhe40^−/−), red pulp macrophages (pooled from 3 experiments, n=9 Bhlhe40^+/+, 8 Bhlhe40^−/−), Kupffer cells, and kidney macrophages (both pooled from 2 experiments, n=10/group) from Bhlhe40^+/+ and Bhlhe40^−/− mice. d, Flow cytometry of Ki67 expression by Bhlhe40^+/+ and Bhlhe40^−/− LPMs (representative of 7 experiments, n=24 Bhlhe40^+/+, 22 Bhlhe40^−/−). e, Frequency of Ki67⁺ LPMs as in d, SPMs, and B cells (both pooled from 8 experiments, n=30 Bhlhe40^+/+, 29 Bhlhe40^−/−) from Bhlhe40^+/+ and Bhlhe40^−/− mice. f, Flow cytometry of BrdU incorporation by Bhlhe40^+/+ and Bhlhe40^−/− LPMs (representative of 5 experiments, n=18/group). g, Frequency of BrdU⁺ LPMs as in f. h, Flow cytometry of pHH3 expression by Bhlhe40^+/+ and Bhlhe40^−/− LPMs (representative of 4 experiments, n=12 Bhlhe40^+/+, 11 Bhlhe40^−/−). i, Frequency of pHH3⁺ LPMs as in h. j, Flow cytometry for discrimination of cell cycle phases of Bhlhe40^+/+ and Bhlhe40^−/− LPMs (representative of 4 experiments, n=12 Bhlhe40^+/+, 11 Bhlhe40^−/−). k, Numbers of LPMs in each phase of the cell cycle as in j. Data in c,e,g,i,k are mean ± s.e.m; each symbol represents an individual mouse (c,e,g,i,k). *P ≤ 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001, unpaired two-sided Student’s t-test.