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. 2021 Nov 5;9:772669. doi: 10.3389/fcell.2021.772669

FIGURE 5.

FIGURE 5

PD-L1 is critical for mammary regeneration in transplantation. (A) Illustration of the 3D culture and transplantation assays. Mammary basal cells were isolated by FACS from WT, PD-L1+/– or PD-L1–/– mice for 3D Matrigel culture or transplantation experiments. (B) Colony size of basal cells from WT, PD-L1+/–, or PD-L1–/– mice in Matrigel culture. Data are presented as mean ± SD. Scale bars, 50 μm. (C) Regeneration frequency of WT, PD-L1+/–, or PD-L1–/– basal cells. The mammary outgrowth numbers and sizes (shown as the percentage of fat pad filled) are combined from three independent experiments. The mammary reconstitution unit (MRU) frequency is shown for each group. Student’s t-test: **** P < 0.0001. (D) Representative whole-mount images of outgrowths from transplantation of WT, PD-L1+/–, or PD-L1–/– basal cells. Scale bars, 1 mm. (E) Regeneration efficiency of pWPI or pWPI-PD-L1 primary mammary cells. The mammary outgrowth numbers and sizes (shown as the percentage of fat pad filled) are combined from three independent experiments. The mammary reconstitution unit (MRU) frequency is shown for each group. Student’s t-test: **** P < 0.0001. (F) Representative whole-mount images of outgrowths from transplantation of pWPI or pWPI-PD-L1 primary mammary cells. Scale bars, 0.5 mm. FACS, fluorescence-activated cell sorting; PD-L1, programmed cell death ligand 1.