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. 2017 Sep 4;214(9):2629–2647. doi: 10.1084/jem.20161855

Figure 3.

Figure 3.

M-CSFR expression is reduced after Raptor deletion in developing myeloid precursors. (A) Expression of CD115 on WT and Rptor−/− macrophages after liquid culture of BM cells with M-CSF (10 ng/ml) for 5 d, with mean fluorescence intensity (MFI) plotted within graph. (B) Expression of CD115 on CD11b+ cells in the spleen of WT and Rptor−/− mice. (C) Expression of CD115 on supernatant and adherent cell fractions of WT and Rptor−/− Lin– BM cells after liquid culture with M-CSF (10 ng/ml) for 1, 2, or 3 d, with MFI plotted above graphs. (D) Analysis of Csf1r mRNA in WT and Rptor−/− Lin– BM cells after liquid culture with M-CSF (10 ng/ml) for 0, 1, 2, and 3 d. (E) Expression of CD115 on WT and Rptor−/− myeloid progenitor cell populations, with MFI plotted within graphs. (F) Flow cytometry analysis of CD115 together with PU.1, IRF8, or KLF4 in WT Lin– BM cells. (G) Expression of PU.1, IRF8, or KLF4 in various myeloid progenitor cell populations of WT and Rptor−/− mice, with MFI plotted above graphs. (H) Flow cytometry analysis of PU.1, IRF8, or KLF4 with CD115 in WT Lin– BM cells 2 d after liquid culture with M-CSF (10 ng/ml). (I) Flow cytometry analysis of PU.1, IRF8, or KLF4 in WT Lin– BM cells after liquid culture with M-CSF (10 ng/ml) for 2 d. Right, summary plot of PU.1, IRF8, or KLF4 in Rptor−/− cells with expression relative to WT controls. Data are mean ± SEM and representative of four (A and C), three (B and F–I), two (D), or six (E) independent experiments. *, P < 0.05; **, P < 0.01; NS, not significant; Student’s t test.