Supplementary Fig. 1.

(A) Acute colitis was induced in 8-week-old C57BL/6 mice (20-24 g) with 3% DSS (MP Biomedicals, USA) dissolved in drinking water given ad libitum for 7 days. During the experimental period, clinical signs, body weight and stool consistency were monitored and recorded daily. On day 8, mice were sacrificed and the paraffin sections of colon tissues were stained with hematoxylin and eosin. (B) The protein levels of LC3B, p-mTOR and related molecules in DSS-induced model in C57BL/6 mice were determined by western blot. Data are the representative of three independent experiments with similar results. (C) Graphs shows a quantitative analysis of p62 and LC3B time-dependent expression in Caco-2 and HT-29 cells stimulated with LPS (1 μg/ml). Bars in the graphs represent mean ± s.e.m. Significant differences from vehicle-treated cells are shown by *p < .05, ** p < .01, ***p < .001. (D) In a preliminary dose-response experiment, mice were randomly assigned to four groups (n = 5) by different dose of rapamycin (0, 0.5, 1, 1.25 mg/kg/day) administrated for 10 days, then sacrificed for LC3B expression assessment by western blot. The results showed that 1.25 mg/kg/day rapamycin has the best inductive effects of autophagy activity. In another preliminary dose-response experiment, mice were randomly assigned to five groups (n = 5) by different dose of 3-MA (0, 2.5, 5, 7.5, 10 mg/kg/day) administrated for 10 days, then sacrificed for LC3B expression assessment by western blot. The results showed that 10 mg/kg/day 3-MA has the best inhibition effects of autophagy activity. Then the dose of 1.25 mg/kg/day rapamycin and 10 mg/kg/day 3-MA were chosen for subsequent experiment. Bars in the graphs represent mean ± s.e.m. Significant differences from vehicle-treated mice are shown by *p < .05, ** p < .01, ***p < .001. (E) The knockdown efficiency of TLR4, MyD88, NF-κB p65 and p-ERK inhibitor GDC-0994 in Caco-2 and HT-29 cells were analyzed by western blot. (F) HT-29 cells were pretreated with different pharmacological or genetic treatments before LPS stimulation for another 6 h. NF-κB p65 translocation was determined by immunofluorescence assay. Scale bar = 20 μm. The data shown were from three independent experiments. In all cases, bars in the graphs represent mean ± s.e.m. Statistical analyses were performed by unpaired t-test or one-way ANOVA. Significant differences from cells treated without LPS are shown by *p < .05, ** p < .01, ***p < .001.