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. Author manuscript; available in PMC: 2015 Oct 1.
Published in final edited form as: J Gastroenterol. 2013 Oct 7;49(10):1378–1391. doi: 10.1007/s00535-013-0885-z

Fig. 3. IL6 induces MUC13 expression via binding of transcription factor STAT5 to the promoter of MUC13.

Fig. 3

(A) MUC13 expression levels vary among cell lines: RNA was isolated from colon cancer (SW48, SW480, SW620, T84), ovarian cancer (SKOV-3, CaOV-3) and pancreatic cancer (HPAFII, MiaPaca) cell lines and analyzed by Q-RT-PCR for MUC13 mRNA. β2-microglobulin was used as the housekeeping gene control. For quantitative analysis, the relative MUC13 RNA expression is normalized to SW48 cell line. (B) Expression profile of transcription factors predicted to bind to the MUC13 promoter: RNA isolated from T84 (top) and HPAFII (bottom) cell lines was analyzed by Q-RT-PCR to determine the expression level of the panel of potential transcription factors identified by in silico analysis. Relative mRNA expression of transcription factors were calculated using ΔΔCt method. (C) Schematic representation of STAT5 DNA binding sequence in the predicted promoter of MUC13: Location of the STAT5 binding sequence is boxed and the STAT5 binding sequence is in red and underlined, illustrating the STAT5 binding site is in a 1 kbp upstream region from the translational start site (TSS). (D) STAT5 binds to the predicted promoter of MUC13: ChIP analysis was performed as described in experimental procedures and MUC13 DNA that was pulled down by anti- STAT5 was amplified using semi quantitative PCR in T84 (D, top left) and HPAFII (D, top right) cells. Densitometry analysis of PCR product is shown in T84 (D, bottom left) and HPAFII (D, bottom right). (E) The MUC13 amplification was confirmed by real time PCR in T84 (right) and HPAFII (left).