Macrophages are crucial for epithelial cell death and adipocyte repopulation during mammary gland involution (original) (raw)
Skip Nav Destination
RESEARCH REPORT| 15 January 2012
Supplemental Figure S1Fig. S1. Confirmation of macrophage depletion with AP20187 treatment in Mafia mice during postpartum involution. (A) Flow cytometry (left) of CD45+ cells shows depletion of macrophages (red, F4/80+ GFP+) in peripheral blood, whereas neutrophils (green, F4/80neg GFPlo) remain following AP20187 treatment. Quantification (right) confirms significant decrease in CD45+ F4/80+ GFP+ cells in peripheral blood at all timepoints, *, P<0.007; _n_=3-5. (B) Quantification of average number of acini per lobule. *, _P_=0.0002; _n_=3. (C) H&E stain of mammary tissue 3 days post-weaning from wild-type (WT) C57BL/6J mice treated during postpartum involution with AP20187 shows no difference from vehicle-treated mice, confirming that AP20187 does not interrupt post-weaning mammary regression in WT mice. (D) GFP IHC (left) shows that mammary adipocytes do not express the transgene on InvD3 in vehicle-treated Mafia mice. H&E stain (middle) shows mammary adipocytes are not targeted by AP20187 treatment in nulliparous Mafia mice. H&E stain (right) of abdominal adipocytes harvested from AP20187-treated InvD3 Mafia mice confirms that AP20187 does not systemically target adipocytes during postpartum involution. (E) Smooth muscle actin (SMA) and p63 IHC show that mammary myoepithelial cells are not directly targeted by AP20187 treatment. Further, myoepithelial marker staining patterns in macrophage-depleted glands resemble WT lactational glands. InvD, involution day; Veh, vehicle; MG, mammary gland; Nullip, nulliparous; AA, abdominal adipocytes; LacD, lactation day. Error bars indicate s.e.m. Scale bars: 150 µm in C; 100 µm in D; 60 µm in E.
- pdf file