Fig 1: Upregulation of TUG1, TET2, and Inflammation and Downregulation of miR-200a-3p Under OGD/R Conditions. A: Cell viability was determined by MTT assay after deoxygenation for 0, 2, 4, 6, 8, 12 h, then following by reoxygenation for 24 h. B: TUG1, miR-200a-3p, NLRP3, and TET2 were detected by qRT-PCR after subject to ORG 12 h/R 24 h condition. C: IL-1ß and IL-18 were measured by ELISA after subject to ORG 12 h/R 24 h condition. D: TET2, NLRP3, Caspase-1, GSDMD-N, IL-18, and IL-1ß were determined by western blot after subject to ORG 12 h/R 24 h condition. Four independent experiments were repeated, with three replicates each time. *p < 0.05, **p < 0.01, ***p < 0.001.
Fig 2: Immunohistochemical staining of the expression levels of AMPK and TET2 in CRC specimens. (A-D) AMPK and (F-H) TET2 proteins were mainly detected in the cytoplasm and membrane of CRC cells. Representative staining patterns of CRC with (A and E) negative, (B and F) weak, (C and G) moderate and (D and H) strong staining intensity. Magnification, ×200. CRC, colorectal cancer; AMPK, AMP-activated protein kinase; TET2, ten-eleven translocation 2.
Fig 3: Knockdown of TUG1 Upregulated miR-200a-3p and Decreased TET2 and Inflammation. A: The expressions of TUG1 after TUG1 knockdown were measured. B: miR-200a-3p was detected by qRT-PCR under ORG 12 h/R 24 h condition. C: Cell viabilities were measured by CCK-8 assay in ORG 12 h/R 24 h -induced SH-SY5Y and SK-N-SH cells after transfection with shNC or shTUG1. D: ELISA measured IL-18 and IL-1ß in ORG 12 h/R 24 h -induced cells with or without TUG1 knockdown. E: TET2 and inflammation-related proteins were detected by western blot in ORG 12 h/R 24 h -induced cells with or without TUG1 knockdown. Four independent experiments were repeated, with three replicates each time, *p < 0.05, **p < 0.01, ***p < 0.001.
Fig 4: Kaplan-Meier survival curves of disease-free survival. (A and B) Survival analysis based on (A) AMPK and (B) TET2 expression levels. AMPK, AMP-activated protein kinase; TET2, ten-eleven translocation 2.
Fig 5: Demethylation of TUG1 by TET2 Promoted Inflammatory Response by Regulation of NLRP3. A: TUG1 expression was measured by qRT-PCR in ORG 12 h/R 24 h -injured cells after TET2 knockdown. B: The DNA methylation and expression of TUG1 were analyzed by methylation-specific PCR in ORG 12 h/R 24 h -induced SK-N-SH cells after TET2 knockdown. C: Cell viabilities were measured in ORG 12 h/R 24 h -induced SH-SY5Y and SK-N-SH cells after cotransfection with shTET2 and pcDNA3.1-TUG1. D: IL-18 and IL-1ß were assessed by ELISA in ORG 12 h/R 24 h - injured cells after TET2 knockdown single or together with TUG1 overexpression. E: NLRP3, GSDMD-N, and Caspase-1 levels were assessed by western blot in ORG 12 h/R 24 h -induced injured cells after TET2 knockdown single or together with TUG1 overexpression. Four independent experiments were repeated, with three replicates each time, *p < 0.05, **p < 0.01, ***p < 0.001.
Supplier Page from Abcam for Anti-Tet2 antibody