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. 2015 Nov 30;30(3):1263–1275. doi: 10.1096/fj.15-278606

Figure 4.

Figure 4.

mTOR regulates differentiated IECs through both mTORC1 and mTORC2. A) Western blot shows loss of mTOR signaling components and downstream signaling activity in WT, RptorΔIEC, RictorΔIEC, and double RptorRictorΔIEC ileal crypts (n = 2 mice). B) Representative H&E, LYZ (brown), and hematoxylin (blue) IHC staining for WT, RptorΔIEC, RictorΔIEC, and double RptorRictorΔIEC ileal tissue. C) Representative RictorΔIEC, RptorΔIEC, and RptorRictorΔIEC ileal tissue stained for alkaline phosphatase (red), Muc2 (green), or ChgA (green) and counterstained with DAPI, DNA (gray), or E-cadherin (magenta). D) Average MUC2 area in WT vs. RictorΔIEC, RptorΔIEC, or RptorRictorΔIEC (dual) ileal goblet cells. E) Average percentage of MUC2+ IECs in RictorΔIEC, RptorΔIEC, or RptorRictorΔIEC (dual) ileal epithelium. F) Average percentage of ChgA+ IECs in WT vs. RictorΔIEC, RptorΔIEC, or RptorRictorΔIEC (dual) ileal epithelium. D–F) Dotted lines indicate levels of shared WT controls shown in Fig. 2C, D, F for ease of comparison. G) Average mitotic index (PHH3, M-phase marker) of WT vs. RptorΔIEC, RictorΔIEC, and RptorRictorΔIEC mice ileal crypts. H) Western blot demonstrates AKT activation in WT and mTORΔIEC and in WT, RptorΔIEC, RictorΔIEC, and RptorRictorΔIEC isolated ileal crypts (from same Western blot as shown in Figs. 1C and 4A, respectively). I) Representative Western blot of WT enteroids treated with the mTOR kinase inhibitor AZD8055 at 50–200 nM for 6 h. V, vehicle. n = 2 independent experiments. B–G) n = 3 mice each genotype. D–G) Error bars represent sd. *P < 0.05; **P < 0.01. Scale bars, 100 μm.