FIG. 4.

Properties of monocyte-derived ATMs. Differentiation of injected monocytes into inflammatory ATMs was analyzed as CD11c⁺ vs. CD11c⁻ ATMs in NC/lean (A) and HFD-fed/obese (B) mice at the indicated times and then plotted as the mean ± SEM from three independent experiments. n = ~8–9 in each group. C: The increase in triply positive cells accounts for the majority of the increase in total ATMs. The numbers of cells of total ATMs (CD11b⁺F4/80⁺), doubly positive (F4/80⁺CD11b⁺CD11c^-), and triply positive (F4/80⁺CD11b⁺CD11c⁺) cells were calculated from FACS data and SVF cell counts and expressed per gram of eWAT. n = 6 per group. *P < 0.05 vs. NC. Data are means ± SEM. D: To detect proliferation of PKH26-labeled monocytes, the proliferation marker BrdU was injected twice at 24 and 3 h before the end point experiment at day 7 after PKH26⁺ cell injection. PKH26⁺ and PKH26⁺ BrdU⁺ ATMs were analyzed by a FACS (left) and then plotted as the mean ± SEM from three independent experiments (right). n = 6. The scattergram is representative of five to six independent mice. E: Immunohistochemistry analyses of the proliferation marker, Ki67, to detect proliferation of PKH26⁺ ATMs in adipose tissue. The image is representative of similar results from three to four independent experiments. Scale bar represents 100 μm. (A high-quality digital representation of this figure is available in the online issue.)