Fig 2: Basal expression levels of COX-1 and COX-2 in Het-1A and KYSE-270 cells. (a) The basal gene expression levels of cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) in KYSE-270 cells were higher than those in Het-1A cells. (b) The basal level of COX-2 protein expression was substantially higher in KYSE-270 cells than in Het-1A cells. The shown blots were cropped to improve the conciseness and the full-length blots are presented in Supplementary Fig. S6. (c) Treatment with the COX-2-selective inhibitor NS-398 (0.2 µmol/L) significantly inhibited PGE2 production in KYSE-270 cells treated with pH 4.5 medium or with 400 µmol/L CDCA. Data are presented as means ± SD (n = 3). PGE2 prostaglandin E2; CDCA chenodeoxycholic acid. Statistical significance was determined by Student’s or Aspin–Welch's t-test or Tukey–Kramer test; *P < .01, †P < .001.

Fig 3: Effects of AGR on (A) protein production, (B) mRNA expression of iNOS, COX-2, and (C,D) production of HO-1, Nrf-2. Microglia cells were stimulated with LPS for (A,B) 12 h, (C) 6 h (HO-1) or 3 h (Nrf-2), or (D) 3 h. The histograms show protein and mRNA expression levels relative to those of ß-actin or TBP. Data represent the mean ± SEM of three independent experiments. * p < 0.05, ** p < 0.001, and *** p < 0.0001 were calculated from comparisons with LPS stimulation values.

Fig 4: 5-MTP blocks PMA-induced p300 HAT activity in SF-Fb. A). SF-Fb were pretreated with 5-MTP at indicated concentrations for 30 min followed by PMA for 4 h. p300 HAT activity was measured. The error bars show mean ± SEM (n = 3). B). SF-Fb were treated with 5-MTP (10 µM) in the absence of PMA (left panel) or presence of PMA (right panel). Right panel also shows treatment of pFb with or without 5-MTP followed by PMA. Each error bar refers to mean ± SEM (n = 3). “NS” denotes statistically non-significant. C). SF-Fb were pretreated with 5-MTP (10 µM) for 30 min followed by PMA (100 nM) for 4 h. COX-2 proteins were analyzed with Western blotting. This Western blot is representative of two experiments with similar results. D). SF-Fb or pFb were treated with or without PMA (100 nM) for 4 h. p300 proteins were isolated by IP, washed and treated with 5-MTP (10 µM) for 30 min. HAT activity was measured. The error bars refer to mean ± SEM (n = 3). “NS” denotes statistically non-significant.

Fig 5: NZD suppresses LPS-stimulated inflammatory response in RAW264.7 Cells. RAW264.7 cells were pretreated with NZD for 1 h and then stimulated with LPS (1 µg/mL) for 18 h. (A) The level of nitrite was determined by Griess reagent. (B, C) RAW264.7 cells were pretreated with NZD for 1 h and then stimulated with LPS (1 µg/mL) for 8 h. The level of NO was measured by a flow cytometer with a fluorescence probe DAF-FM. (D, E, F) The expressions of iNOS and COX-2 were detected by Western blotting. * p < 0.05, ** p <0.01, ***p < 0.001 vs. LPS group.