lncRNA TUG1 Facilitates Colorectal Cancer Stem Cell Characteristics and Chemoresistance by Enhancing GATA6 Protein Stability

Figure 3

Analysis of the interaction between lncRNA TUG1 and GATA6. (a) RNA Interactome Database and COAD colon cancer database were applied to predict the transcription factors that had potential interactions with lncRNA TUG1 and were highly expressed in CRC. (b) RNA pull-down assay was conducted to verify the interaction between lncRNA TUG1 and ETV4, ASCL2, GATA6, and SOX9. (c) Verification of the interaction between lncRNA TUG1 and GATA6 by RNA immunoprecipitation assay. (d, e) si-TUG1 was transfected into CD133^-/CD44^- or CD133⁺/CD44⁺ isolated from SW480 cells. The mRNA and protein levels of GATA6 were analyzed using qRT-PCR and Western blot. (f, g) si-TUG1 was transfected into CD133^-/CD44^- or CD133⁺/CD44⁺ isolated from SW480 cells, and the cells were treated with 100 μg/ml cycloheximide (CHX) for 0, 4, and 8 h. Detection of the GATA6 protein level by Western blot. (h–k) si-TUG1 was transfected into CD133^-/CD44^- or CD133⁺/CD44⁺ isolated from SW480 cells. The mRNA and protein levels of BMP4 and BMP2 were assessed using qRT-PCR and Western blot. vs. si-NC or CD133^-/CD44^-. vs. IgG or CD133^-/CD44^-.

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