Journal: The Journal of Biological Chemistry

Article Title: Pentacyclic triterpenes modulate farnesoid X receptor expression in colonic epithelial cells: Implications for colonic secretory function

doi: 10.1016/j.jbc.2022.102569

Figure Lengend Snippet: Hederagenin is not an agonist of FXR. T 84 cells were serum-starved for 24 h and bilaterally treated with DMSO (1%), GW4064 (5 μM), or HG (5–100 μM) for 24 h. A , basolateral supernatants were collected, and FGF-19 protein was analyzed by ELISA (n = 4; ∗ p < 0.05). B , FGF-19 mRNA expression was analyzed by qRT-PCR (n = 3). C , TEER was assessed pretreatment and posttreatment (n = 7; ∗∗∗ p < 0.001). D , FXR reporter cells were treated with DMSO (0.1%), GW4064 (5 μM), or HG (1–10 μM) for 24 h. FXR activation was assessed by luminescence measurement (n = 3; ∗∗ p < 0.01). Data are expressed as mean ± SD. Statistical analysis was performed using either repeated measures one-way ANOVA with Dunnett’s post hoc test or two-way ANOVA with Sidak’s post hoc test, as appropriate. FXR, farnesoid X receptor; HG, hederagenin; qRT-PCR, quantitative real-time PCR.

Article Snippet: An antibody pair was used for human FGF-19 from DuoSet ELISA Development System from R&D Systems (Catalog # DY969), and procedure was carried out as per manufacturer’s protocol.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Activation Assay, Real-time Polymerase Chain Reaction

Journal: The Journal of Biological Chemistry

Article Title: Pentacyclic triterpenes modulate farnesoid X receptor expression in colonic epithelial cells: Implications for colonic secretory function

doi: 10.1016/j.jbc.2022.102569

Figure Lengend Snippet: Pentacyclic triterpenes potentiate agonist-induced FXR signaling in T 84 colonic epithelial cells. T 84 cells were serum-starved for 24 h and bilaterally pretreated with HG (5 μM) for 1 h prior to 24 h treatment with varying concentrations of GW4064 (0.5–5 μM). A , FGF-19 mRNA expression was analyzed by qRT-PCR (n = 8; ∗∗ p < 0.01, ∗ p < 0.05, # p < 0.05). B , basolateral supernatants were collected and FGF-19 protein expression was assessed by ELISA (n = 9, ∗ p < 0.05). C , T 84 cells were bilaterally pretreated with HG (5 μM) for 1 h prior to 24 h treatment with CDCA (50 μM). Basolateral supernatants were collected, and FGF-19 protein expression was assessed by ELISA (n = 8, ∗ p < 0.05, ∗∗ p < 0.01, # p < 0.05). D , T 84 cells were bilaterally pretreated with OA (5 μM) for 1 h prior to 24 h treatment with GW4064 (5 μM). Basolateral supernatants were collected, and FGF-19 protein expression was assessed by ELISA (n = 4, ∗ p < 0.05, # p < 0.05). Data are expressed as mean ± SD. Statistical analysis was performed using repeated measures one-way ANOVA with Tukey’s post hoc test. ∗denotes significant differences compared to untreated controls, and # denotes significant differences compared to GW4064-treated cells. FXR, farnesoid X receptor; HG, hederagenin; OA, oleanolic acid; qRT-PCR, quantitative real-time PCR.

Article Snippet: An antibody pair was used for human FGF-19 from DuoSet ELISA Development System from R&D Systems (Catalog # DY969), and procedure was carried out as per manufacturer’s protocol.

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction