Fig 2: Tiep2-deficient mice were more resistant to AOM/DSS-induced tumorigenesis.A The protocol of AOM/DSS-induced CRC. The tumor number was significant lower in Tipe2-deficent AOM/DSS models (B), especially tumors less than 2 mm in diameter (C). Tipe2-defiency showed less severe intestinal morphology (D) accompanied by longer length of the colon (E) in AOM/DSS-induced mice. Tipe2-deficent AOM/DSS models showed lower serum levels of proinflammatory cytokines, such as IL-6 (F), MCP-1 (G), IL-12 (H) and TNF-a (I). Data are representative of three independent experiments and expressed as means ± SEM. Significant difference between two groups was determined using an unpaired two-tailed Student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001.

Fig 3: Src controls H3 hyperacetylation upon LPS/IFN? stimulation through p300.iNOS expression (a), MCP-1 production (b), and Nos2 and Ccl2 gene expression levels (c) at 16 h post 1 ng/ml LPS/IFN? stimulation in siSrc-treated cells portray Src as an inflammatory gene. Histone acetylation staining in cells treated with siSrc for 48 h (d), and a further 16 h of 1 ng/ml LPS/IFN? stimulation (e) show lower global H3Ac levels with Src knockdown. (f) Total HAT and p300 HAT activity measured in siSrc-treated cells 2 h post LPS/IFN? stimulation show lower p300 HAT activity. Data has been represented as mean ± SEM, and one-way ANOVA with multiple comparisons using Tukey test was performed. Exceptions in panels b–e where only 2 conditions are available were instead subjected to Student’s t-test for analysis. Violin plots show quartiles and median. Source data are provided.

Fig 4: Myeloid-specific GP96 deficiency prevents chronic alcohol-induced endotoxin and pro-inflammatory cytokine production. (A) Endotoxin level was measured in serum (n = 8-12). (B) CyP2e1 was detected in liver microsomal fraction by western blot, and calnexin was used as an internal loading control (n = 3). Total RNA from liver tissue was subjected to quantitative RT-PCR for analysis of pro-inflammatory cytokines TNF-a, IL-6, MCP-1, and IL-1ß; NLRP3; anti-inflammatory markers IL-10, TGF-ß, and ATF3 (C); and macrophage markers (n = 6-10) (F). (D) Total liver protein level for TNF-a was measured in tissue extracts of pair-fed and alcohol-fed mice by ELISA (n = 6-10). (E) ATF3 protein level was analyzed by western blot in whole-liver extracts using tubulin as an internal control (n = 3). The mRNA expression profile of (G) pro-inflammatory, and (H) anti-inflammatory and restorative macrophage markers, Trem2 and ATF3, were analyzed in liver macrophages of pair-fed and alcohol-fed mice (n = 5). (I) The mRNA level of pro-inflammatory cytokines was analyzed in BMDMs isolated from WT and M-GP96KO mice and stimulated with 100 ng/mL LPS for 2 hours (n = 9). Data are represented as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Fig 5: Src activation drives H3 hyperacetylation through p300.(a) Effects of PP1 (5 µM) and PP2 (5 µM) treatment on Src activation measured by Western blot. Anti-inflammatory effects of PP1 and PP2 treatments demonstrated by MCP-1 production (b), Ccl2 and Nos2 gene levels (c), and iNOS staining (d) in cells treated with Src inhibitors for 6 h, followed by stimulation of 1 ng/ml LPS/IFN? for 16 h. Global H3Ac levels in cells treated with PP1 and PP2 for 24 h (e), and 6 h Src inhibitors followed by 16 h 1 ng/ml LPS/IFN? treatment (f) show Src inhibition suppress H3Ac expression. (g) p300 activity in unstimulated and stimulated cells wherein 6 h drug treatment was followed by 2 h of 1 ng/ml LPS/IFN? treatment show suppressed p300 activity in PP1- and PP2-treated cells. (h) Ccl2 proximal promoter H3Ac enrichment measured at two different loci in 6 h PP1- and PP2-treated cells that were subsequently stimulated for 2 h with 1 ng/ml LPS/IFN?; ChIP-qPCR with pre-immune IgG displayed background signal and was comparable across conditions (data not shown). (i) Src proximal promoter H3Ac enrichment measured in 6 h PP1- and PP2-treated cells that were subsequently stimulated for 2 h with 1 ng/ml LPS/IFN?. (j) Src gene expression in in 6 h PP1- and PP2-treated cells that were subsequently stimulated for 16 h with 1 ng/ml LPS/IFN?. Data has been represented as mean ± SEM, and one-way ANOVA with multiple comparisons using Tukey test was performed. Violin plots show quartiles and median. Source data and unprocessed blots are provided.