Fig. 3

Fig. 3

a qRT-PCR analysis of GLI1 mRNA expression in HT-29 and Caco-2 cells with or without LTC₄ stimulation for 48 h. b Western blot analysis of 15-PGDH, GLI1, and phospho-PKA (αβγ subunit and β subunit) levels in HT-29 and Caco-2 cells with or without LTC₄ stimulation. α-Tubulin served as the loading control. c Immunofluorescence analysis of GLI1 in HT-29 cells with or without LTC₄ stimulation for 48 h. d qRT-PCR analysis of HT-29 cells transfected with control shRNA (shCTRL) or PGDH-specific shRNA (shHPGD) with or without LTC₄ stimulation for 48 h. e Western blot analysis of HT-29 cells transfected with control shRNA (shCTRL) or PGDH-specific shRNA (shHPGD) blotted with antibodies against 15-PGDH, GLI1, or phospho-PKA (αβγ subunit and β subunit) with or without LTC₄ stimulation for 48 h. α-Tubulin served as the loading control. f Immunofluorescence analysis of GLI1 in HT-29 cells transfected with control shRNA (shCTRL) or PGDH-specific shRNA (shHPGD) with or without LTC₄ stimulation for 48 h. g qRT-PCR analysis of 15-PGDH and GLI1 in HT-29 cells treated with the PKA inhibitor H89 (PKA-inh) for 6 h followed by LTC₄ for 48 h. h Western blot analysis showing the expression of 15-PGDH, GLI1, and phospho-PKA (αβγ subunit and β subunit) in HT-29 cells treated with the PKA inhibitor H89 (PKA-inh) for 6 h followed by LTC₄ for 48 h. i Immunofluorescence analysis of GLI1 in HT-29 cells treated with the PKA inhibitor H89 (PKA-inh) for 6 h followed by LTC₄ for 48 h. HPRT1 was used as the housekeeping gene for normalisation of the qRT-PCR gene expression data. Graphs represent the mean ± SEM of data from 3 to 4 independent experiments, **P < 0.01, ***P < 0.001.