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. Author manuscript; available in PMC: 2022 Jan 18.
Published in final edited form as: Cell Rep. 2020 Aug 18;32(7):108049. doi: 10.1016/j.celrep.2020.108049

Figure 4. 15-LOX-1 Downregulates LRP5 Protein Expression via 13-HODE.

Figure 4.

(A) Eicosanoid metabolite profile of the IECs from ApcΔ580 and ApcΔ580-15-LOX-1 mice, as described in Figures 1D–1F, measured using liquid chromatography-tandem mass spectrometry (LC-MS/MS) (n = 14 mice per group). Values are mean ± SEM.

(B–E) Spearman correlation analysis of colonic tumor volumes and levels of 13-HODE (B), 15-HETE (C), LXA4 (D), and LXB4 (E) in IECs from the mice as described in (A) (n = 56 for all four groups).

(F and G) Cell viability of colonic organoids derived from the IECs of ApcΔ580 mice treated with 13(S)-HODE at 0, 3, 6, or 13.5 μM (F) or treated with 13(S)-HODE (3 μM), 15(S)-HETE (2.15 μM), LXA4 (8.86 nM), or LXB4 (36.24 nM) for 6 days (G), measured using CellTiter-Glo Luminescent Cell Viability Assay (n = 3 repeats). Values are mean ± SEM. *p < 0.05 and **p < 0.01 (one-way ANOVA).

(H) LRP5 and active β-catenin protein expression levels in organoid cells treated with 13(S)-HODE at the indicated concentrations for 6 days, as described in (F).

(I and J) Levels of 13-HODE in SW480 (I) and LoVo (J) cells stably transduced with either control (Ctrl) or 15-LOX-1 lentivirus, measured using LC-MS/MS in triplicated measurements. Values are mean ± SD. ***p < 0.001 (unpaired t test).

(K) 15-LOX-1, LRP5, and active β-catenin protein levels in SW480 and LoVo cells transduced with either Ctrl or 15-LOX-1 lentivirus, measured using western blot analysis.

(L) LRP5 and active β-catenin protein levels measured using western blot analysis in SW480 and LoVo cells supplemented with 13(S)-HODE at the indicated concentrations in cell culture media for 48 h.