Fig 2: The expression of DLG2 in colon cancer and the effect of low DLG2 level in colon cancer cells. a The expression of DLG2 in 20 paired samples from the healthy ascending colon (Healthy A), healthy descending colon (Healthy D), mucosa from colon cancer patients 10 cm adjacent to the tumor (Mucosa) and colon cancer tumor (Tumor). b TheDLG2 expression in colon adenomas comparing 32 samples < 1 cm, 16 samples between 1 and 1.5 cm and 15 samples larger than 1.5 cm. c The effect of DLG2 silencing (siDLG2) and overexpression (DLG2) on SW480 cell proliferation 48 h post transfection. d Kaplan–Meier survival curve showing the 5-year event free survival probability of colon cancer patients. e–g The gene expression of e NLRP3, f NFKBIZ, and g FOXO3 in response to DLG2 overexpression in SW480 cells. Normalized level of protein phosphorylation of h AKT1, i FOXO3 and j S6 in response to DLG2 overexpression in SW480 cells. k Representative immunoblot showing the effect of DLG2 overexpression on SW480 cells showing the expression of DLG2, p-AKT (s473), AKT1, p-FOXO3a (s318/321), FOXO3, p-S6 (s235/236) and S6, normalized to GAPDH. Each experiment was performed in triplicate. The expression data are presented as log2 fold change and plotted as either Tukey’s box and whisker plots showing IQR, line at the median, + at the mean with whiskers ± 1.5-fold of interquartile range or as mean. *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 response of NFκβ members to increased DLG2 expression and activation of the inflammasome in THP-1 cells. The expression of a RELA, b RELB, c NFKB1 and d NFKBIZ 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; DLG2, p-P65 (Ser 536), RELA, RELB, p-P105 (Ser 932), NFκB1, IκBζ and GAPDH. Each experiment was performed in triplicate. The gene expression data are presented as log2 fold change and plotted as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001