Fig 1: ART1 knockdown decreased IL-6-induced cell proliferation. (A) Proliferation rates of WT, NC and ART1-sh CT26 cells after incubation with different concentrations of IL-6 (0, 5, 10, 20, 50 and 100 ng/ml) for 48 h. (B and C) Protein expression levels of ART1 in CT26 and LoVo cells transfected with ART1-sh. (D and E) Protein expression levels of c-Myc, cyclin D1 and Bcl-xL in WT, NC and ART1-sh CT26 cells after induction with IL-6 (25 ng/ml) for 24 h. (F and H) DNA synthesis in WT, NC and ART1-sh CT26 cells after incubation with IL-6 (25 ng/ml) for 24 h. Red fluorescence represents DNA synthesis, and blue fluorescence represents the nuclei. The scale bar is 50 μm. (G and I) Colony formation rate of WT, NC, and ART1-sh CT26 cells after incubation with IL-6 (25 ng/ml) for 2 weeks. *P < 0.05, **P < 0.01. WT, wild-type; sh, short hairpin RNA; NC, negative control; ART1, mono-ADP-ribosyltransferase-1
Fig 2: Expression levels of ART1 and gp130 in colorectal cancer tumour and control tissues. The scale bars are 50 and 100 μm. gp130, glycoprotein 130; ART1, mono-ADP-ribosyltransferase-1.
Fig 3: ART1 knockdown inhibited IL-6-induced gp130 and p-STAT3 expression. (A and C) Protein expression level of gp130 and ratio of p-STAT3/STAT3 in NC and ART1-sh CT26 cells after induction with IL-6 (25 ng/ml) for 0.5 or 24 h. (B and D) Protein expression level of gp130 and the p-STAT3/STAT3 ratio in NC and ART1-sh LoVo cells after induction with IL-6 (20 ng/ml) for 0.5 or 24 h. *P < 0.05, **P < 0.01, ***P < 0.001. WT, wild-type; p-, phosphorylated; gp130, glycoprotein 130; sh, short hairpin RNA; NC, negative control; ART1, mono-ADP-ribosyltransferase-1.
Fig 4: Double immunofluorescence staining of ART1 and gp130. (A and B) Double immunofluorescence staining of ART1 and gp130 in WT CT26, LoVo and HCT116 cells. Green fluorescence represents ART1, red fluorescence represents gp130, and blue fluorescence represents the nuclei. The scale bar is 25 μm. The Pearson correlation coefficients were 0.588, 0.632 and 0.782, respectively. WT, wild-type; gp130, glycoprotein 130; ART1, mono-ADP-ribosyltransferase-1.
Fig 5: ART1 knockdown inhibits tumour growth in a colitis mouse model. (A) H&E staining of normal and colitis mouse colon tissues. The scale bar represents 50 μm. (B and C) Volume of transplanted tumours formed in colitis model mice by NC and ART1-sh CT26 cells. (D and E) Protein expression levels of gp130, c-Myc, cyclin D1, and Bcl-xL and the ratio of p-STAT3/STAT3 in the transplanted tumours formed by NC and ART1-sh CT26 cells in colitis model mice. **P < 0.01, ***P < 0.001. p-, phosphorylated; gp130, glycoprotein 130; sh, short hairpin RNA; NC, negative control; ART1, mono-ADP-ribosyltransferase-1.
Supplier Page from Abcam for Anti-ART1 antibody