Fig. 4

Fig. 4

qRT-PCR validation of gene expression in shCTRL (control shRNA)- or shGLI1 (GLI1-specific shRNA)-transfected HT-29 cells with or without LTC₄ stimulation for 48 h. Marker of tumour suppression in a15-PGDH. Markers of differentiation in bSI, in cMUCIN-2, and in dCDHR2. Marker of differentiation regulation as in eCDX2. Markers of Wnt activation in fAXIN2 and gcMYC. Marker of proliferation in h (CCND1). HPRT1 was used as the housekeeping gene for normalization of the gene expression data. i Western blot analysis of whole-cell lysates for 15-PGDH, GLI1, SI, CDX2, and CDHR2 expression in shCTRL (control shRNA)- or shGLI1 (GLI1-specific shRNA)-transfected HT-29 cells with or without LTC₄ stimulation for 48 h. α-Tubulin served as the loading control. j Immunofluorescence analysis of GLI1 and Mucin-2 in cells with or without LTC₄ stimulation for 48 h and transfected with shCTRL or shGLI1. Graphs represent the mean ± SEM of data from three independent experiments, *P < 0.05, **P < 0.01, ***P < 0.001.