Figure 4
Detection of proliferating BAs in the neonatal mice. (A) Representative immunofluorescence images of iBAT. Bar = 25 µm. P8: postnatal day 8. pH3: phospho-Histone H3 (Ser10). (B) Quantification of pH3 positive brown adipocytes at postnatal day 4 and day 8. Student t test, *P < 0.05. Each datum point indicates one individual animal. (C) Experimental design for FACS analysis. In the Ucp1::Cre; Rosa 26tdTom mice, BAs were genetically labeled with tdTomato (tdTom) fluorescence protein. BAs from dissociated iBAT were used for FACS analysis. (D) Representative images of dissociated BAs. Bar = 50 µm. (E,F) FACS analysis of BAs. iBAT from P8 Ucp1::Cre; Rosa 26tdTom mice were dissociated and stained with Hochest. E. tdTom + BAs gating image. (F) DNA content histogram of gated tdTom + BAs. tdTom + BAs (Population 1, P1) were separated into three populations based on DNA contents: P2, P3, P4, representing BAs at G0/G1, S and G2/M phases, respectively. The cell number of P2, P3, and P4 was normalized to the number of gated TdTom + BAs (P1), and calculated percentages were shown in the inset bar graph. N = 4.
