Fig. 4

IL-33/ST2 upregulates COX2 expression through NF-κB signaling. a,b The COX2 mRNA (a) and protein (b) expression in primary CRC cells or HT29 cells responding to the incubation with rhIL33 (100 ng/mL) or/ and ST2 antibody (2 μg/mL) for 24 h. Each experiment was performed three times. Data expressed as mean ± SEM. ** P < 0.01. c ST2 expression distribution in primary CRC cells, sorted ST2-negative and sorted ST2-positive primary CRC cells. The proportion of ST2 positive subset is shown. d Relative COX2 mRNA levels in ST2-negative or ST2-positive primary CRC cells responding to IL-33 (100 ng/mL) incubation for 24 h in 24-well plates (1 × 10⁵ cells per well). Three parallel wells were set for each treatment. Data expressed as mean ± SEM. ** P < 0.01. e The correlation between COX2 and ST2 transcripts in 394 CRC samples recorded in TCGA database. These two sets of data both have a normal distribution. Pearson r = 0.356, P < 0.01. f COX2 mRNA levels in primary CRC cells, HT29 cells and MC38 responding to the incubation with IL-33 (100 ng/mL) or/ and BAY11–7082 (10 μM). Three parallel wells were set for each treatment. Each experiment was performed three times. Data expressed as mean ± SEM. * P < 0.05. ** P < 0.01. g COX2 protein levels in primary CRC cells, HT29 cells and MC38 responding to the incubation with IL-33 (100 ng/mL) or/ and BAY11–7082 (10 μM). Each experiment was performed three times. h The knocking-down efficiency of NF-κB P65 in HT29 cells. The P65 mRNA (left panel) and protein (right panel) were both detected. Data expressed as mean ± SEM. ** P < 0.01. i COX2 mRNA (left panel) and protein (right panel) levels responding to IL-33 incubation (100 ng/mL) for 24 h in HT29 cells transfected with short hairpin RNA expressing plasmid against NF-κB P65 (shP65) or nonsense RNA expressing plasmid (shNC). Each experiment was performed three times. Data expressed as mean ± SEM. ** P < 0.01