Figure 5.

LINC00160 regulates TFF3 via C/EBPβ. A, C/EBPβ is a putative TF regulated by LINC00160, and TFF3 is the downstream target gene of C/EBPβ in the LncMAP database. B, C/EBPβ proteins were pulled down by biotinylated RNA fragments of LINC00160 by the Western blot analysis in contrast to antisense oligos. C, Enrichment of LINC00160 on the C/EBPβ promoter region was detected by RIP‐qPCR assay. D, Three binding sites of C/EBPβ and TFF3 obtained from the JASPAR website. E, The Luciferase activity of pmirGLO‐TFF3 with all three C/EBPβ binding sites or pmirGLO‐TFF3, respectively, truncated three C/EBPβ binding sites was determined in the presence of C/EBPβ overexpressing vectors compared with empty vectors. F, The Luciferase activity of pmirGLO‐TFF3, respectively, mutated at three C/EBPβ binding sites was determined in the presence of C/EBPβ overexpressing vectors compared with empty vectors. G, TFF3 was immunoprecipitated in the presence of C/EBPβ antibody relative to IgG by ChIP assays. H‐J, The mRNA expression and protein level of TFF3 in BC tissue and paired adjacent normal tissue (H), in MCF10A cells, parental MCF‐7 cells, MCF‐7/Tax, parental BT474 and BT474/Dox cells (I), and in LINC00160‐depleted MCF‐7/Tax and BT474/Dox cells compared with scramble siRNA‐treated cells were determined by RT‐qPCR and Western blot analyses. J, The mRNA expression and protein level of TFF3. Data are expressed as mean ± s.d., representative of three independent experiments. One‐way ANOVA was used to determine statistical significance. *P < .05