Fig 2: The response of cytokines and apoptotic proteins to increased DLG2 level and formation of the inflammasome in THP-1 cells. The gene expression of a IL1B, b IL6, c BAX and d BCL2 in response to control conditions (circle symbol), LPS priming (square symbol) and LPS + ATP treatment (triangle symbol) with or without DLG2 overexpression. e Representative immunoblot showing the expression of; BAX, BCL2, p-STAT3, STAT3, p-S6, S6, TIG1 and GAPDH. f Activation of the inflammasome in DLG2 silenced, control and DLG2 overexpressed THP1-ASC-GFP cells, in response to LPS and ATP treatment, as determined by PYCARD/ASC speck formation, presented as a percentage of observed cells. g The transfection efficiency of DLG2 in THP1 cells for silenced, control and DLG2 overexpression. Each experiment was performed in triplicate. The expression data are presented as log2 fold change and plotted as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001

Fig 3: The response of THP-1 cytokine production in colon cancer COLO205 cells. a Representative western blot showing the level of IL-1β and IL-6 production and release in the cell medium in DLG2 silenced, control and DLG2 overexpressed THP-1 cells in response to LPS and ATP, normalized to total protein level. b The effect of cell medium from DLG2 silenced (siDLG2), control and DLG2 overexpressed (DLG2) THP-1 cells on COLO205 cell proliferation, and c percentage of COLO205 cells in G2/M phase. Representative immunoblot showing the effect of THP-1 inflammasome growth media on the expression of d RELA, RELB, IκBζ, NFκB1, e BAX, BCL2, p-STAT3, STAT3, normalized to GAPDH. Each experiment was performed in triplicate. The expression data are presented as log2 fold change and plotted as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001

Fig 4: The response of DLG2, NLRP3 and NFKBIZ gene expression to inflammation in mouse, cell and fly models. a DLG2, e NLRP3 and h NFKBIZ gene expression in the colon tissue of 5 mice for each time point in response to DSS treatment at 0, 2-, 4- and 6-days post treatment (cohort GSE22307). b DLG2, f NLRP3, and i NFKBIZ expression in the colon tissue of 4 mice in response to T-cell transfer at 0, 2-, 4- and 6- weeks post transfer (cohort GSE27302). c DLG2, g NLRP3 and j NFKBIZ expression in THP1 cells in response to LPS treatment at 0, 12 and 24 h. d The expression of dmDLG in Drosophila melanogaster gut in response to lactate bacteria treatment at 0, 1-, 2-, 3-, 4- days post treatment. The expression data are presented as median centered log2 fold change and plotted as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001

Fig 5: The alteration of DLG2, NLRP3 and NFKBIZ gene expression in colon inflammation and cancer. a DLG2, e NLRP3 and i NFKBIZ gene expression by sample type including; 8 Healthy patients, 15 IBD patients, 15 adenoma patients and 15 colon cancer patients (cohort GSE4183). b DLG2, f NLRP3, and j NFKBIZ expression in 20 healthy individuals compared to 206 with Ulcerative colitis (UC) (cohort GSE109142). c DLG2, g NLRP3 and k NFKBIZ expression in 73 patients UC patients with active disease compared to 23 patients with inactive disease (cohort GSE75214). d DLG2, h NLRP3 and l NFKBIZ expression of 24 case-controlled tumor mucosa samples (cohort GSE10950). The expression data are presented as median centered log2 fold change and plotted as Tukeys box and whisker plots showing IQR, line at the median, + at the mean with whiskers ± 1.5-fold of interquartile range. Data outside the whiskers are shown as outliers. The paired data are shown as an individual symbol with a connecting line. *p < 0.05, **p < 0.01, ***p < 0.001