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. 2014 Jul 29;289(37):25737–25749. doi: 10.1074/jbc.M114.570838

FIGURE 3.

FIGURE 3.

MERTK drives efferocytosis in PMECs and when stably expressed in MCF10A cells. A, primary mammary epithelial cells (PMECs) isolated from wild type (WT) or knock-out Mertk(−/−) mice were labeled with CFDA-green. Cells were starved for 6 h and then co-cultured with PKH26-red apoptotic CEM for 5 h and efferocytosis was analyzed by flow cytometry. The experiment was done in triplicate and the representative of 2 independent experiments is shown (n = 2). B, PKH26-red MCF10A-MERTK and MCF10A-pMSCV stable cells were starved for 6 h and then co-cultured with PKH67-green apoptotic CEM in 10% FBS medium for 3 h, and efferocytosis was determined by flow cytometry. C, efferocytosis was carried out as described in B with human GAS6 conditioned medium, GAS6 (1) or (0.5), instead of 10% serum. GAS6 (0.5) was generated by mixing equal volumes of GAS6 (1) with fresh RPMI. NS = not significant. D, representative confocal imaging of PKH26-red MCF10A-MERTK or MCF10A-pMSCV phagocytes and PKH67-green apoptotic CEM co-cultures. The experiment was performed as described in B in an 8-chambered slide.