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. 2014 Jan 13;211(1):15–28. doi: 10.1084/jem.20131080

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© 2014 Li et al.

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Figure 6. — COX-2 expression and prostaglandin production in LAM nodules and LAM patients. (a) Transcript levels of PTGS2 (COX-2), were measured using real-time RT-PCR on RNA prepared from LAM lung and clinically normal lung samples (n = 3 each). Data represent the mean of PTGS2 levels from three subjects from per group. (b) LAM lung tissues stained with COX-2 antibody. Bar, 250 µM. Results are representative of six cases. (c) EBCs were collected from LAM patients before and after aspirin (ASA) treatment (81 mg/day) for 2 wk. Levels of 15-epi-LXA₄ were quantified (n = 3). Data represent the mean of 15-epi-LXA4 levels from three subjects from per group. (d) TSC2-deficient LAM patient-derived 621–101 cells were incubated with 100 nM 15-epi-LXA₄ for 0, 24, 48, and 72 h. Cell proliferation was measured using MTT assay. Results are representative of one out of three experiments including twelve sets of samples in each experiment. (e) The 15-epi-LXA4 integrity was confirmed by UV-Vis spectrophotometry to ensure the presence of the diagnostic tetraene chromophore and accurate quantitation and HPLC to ensure that only a single peak was present without evidence for isomerization. TSC2-deficient 621–101 cells were incubated with 10, 20, 100, 200, or 500 nM 15-epi-LXA₄ for 72 h. Cell proliferation was measured using MTT assay. Results are representative of 1 out of 2 experiments, including 16 sample sets in each experiment. (f) Urine samples were collected from LAM patients and healthy women from two geographic locations. Urinary levels of PGE₂ in LAM (n = 29) and healthy women (N; n = 18) were assessed (ELISA). Serum levels of PGE₂ (g) and 6-keto PGF_1α (h) in LAM (n = 14) and healthy women (N; n = 13) were assessed (ELISA). *, P < 0.05; **, P < 0.01; ***, P < 0.001; Student’s t test for transcript levels and cell growth tests; Mann Whitney test for prostaglandin quantification in clinical data.