Fig 2: Effect of tricellulin knockdown or overexpression on regulation of colorectal cancer HCT116 cell apoptosis. (A) Hoechst 33258 staining. Apoptosis was detected by Hoechst 33258 staining. Magnification, ×200. (B) Western blotting. Expression of the apoptosis markers was detected using western blotting in HCT116 cells after TRIC-KD with or without TNF-a treatment. (C) Western blotting. Expression of the apoptosis markers was assessed using western blotting in HCT116 cells after TRIC-OE with or without PDTC treatment. TRIC-KD, tricellulin knockdown; TRIC-OE, tricellulin overexpression; PDTC, pyrrolidinecarbodithioic acid; CON, control; NC, negative control; TRIC, tricellulin.

Fig 3: Tricellulin expression and association with poor survival of colorectal cancer patients. (A) IHC. Tricellulin expression was significantly higher in colorectal cancer tissues than in normal colorectal mucosae. Magnification, ×400. (B) Immunofluorescence staining of tricellulin in colorectal cancer tissues and normal colorectal mucosae. Magnification, ×400. (C) Summarized data of IHC. The data revealed that 51 (52.0%) colorectal cancer tissues had tricellulin overexpression, whereas 2 (13.3%) normal mucosae had tricellulin expression. (D) Kaplan-Meier curves. Tricellulin expression is associated with poor OS of colorectal cancer patients (n=98). IHC, immunohistochemistry; OS, overall survival; N, normal; Ca, cancerous; TRIC, tricellulin; CRC, colorectal cancer.

Fig 4: Stable knockdown or overexpression of tricellulin in colorectal cancer cells. (A) Western blotting. Tricellulin expression was assessed in a panel of colorectal cancer cell lines and a normal human colon mucosal epithelial cell line NCM460 using western blotting. (B and C) Western blotting and RT-qPCR analysis for TRIC-KD. GAPDH was used as a reference. (D and E) Western blotting and RT-qPCR analysis for TRIC-OE. GAPDH was used as the reference. RT-qPCR, reverse transcription-quantitative PCR; TRIC-KD, tricellulin knockdown; TRIC-OE, tricellulin overexpression; TRIC, tricellulin; CON, control; NC, negative control.

Fig 5: Effects of tricellulin overexpression and knockdown on regulation of colorectal cancer cell invasion and gene expression. (A-C) Transwell assay. Stable HCT116 cells were grown and subjected to Transwell migration and invasion assay (magnification, ×200). The graphs are summarized data of the assays of B and C. (D) FITC-Phalloidin staining. HCT116 cells were grown and stably infected with lentiviruses carrying tricellulin cDNA (TRIC-OE) or shRNA (TRIC-KD) or NC and then subjected to FITC-Phalloidin staining. TRIC-KD, tricellulin knockdown; TRIC-OE, tricellulin overexpression; NC, negative control; TRIC, tricellulin.