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Image Search Results
Journal: Nature Communications
Article Title: SPA70 is a potent antagonist of human pregnane X receptor
doi: 10.1038/s41467-017-00780-5
Figure Lengend Snippet: SPA70 reduces the metabolism and enhances the efficacy of chemotherapeutic agents. a , b SPA70 reduces the metabolism of paclitaxel (PTX) in primary human hepatocytes (PHHs). In a , PHHs were pretreated for 72 h with 0.1% DMSO, 1.6 µM RIF or 0.5 µM or 5 µM PTX in the presence or absence of 10 µM SPA70 before receiving 3.3 µM midazolam (MDZ) and being incubated for a further 4 h. The concentrations of 1′-OH MDZ (an MDZ metabolite) in the medium for each treatment were determined using LC/MS/MS. In b , PHHs were treated with 1 µM PTX in the presence or absence of 10 µM SPA70. The concentrations of PTX metabolites, 3-p-OH PTX (generated by CYP3A4) and 6α-OH PTX (generated by CYP2C8), were determined using LC/MS/MS. Each data point represents the compound concentration measured after 24 or 48 h of treatment. Data are expressed as the mean and 95% confidence interval (CI). c , d hPXR levels inversely correlate with the sensitivity of cancer cells to PTX. In c , LS180 cells with (hPXR overexpression) or without (parental cells) stable exogenous expression of hPXR were treated with PTX in the presence or absence of 10 µM SPA70 for 96 h before a CellTiter-Glo assay was performed. The vehicle was DMSO. In d , SNU-C4 cells treated with pooled siRNAs (siPXR_pool) or individual siRNAs (siPXR_1, 2, 3 and 4) targeting hPXR, or with nontargeting controls (siNT) with or without SPA70, were treated with PTX for 96 h before a CellTiter-Glo assay was performed. The ‘% normalized growth’ was determined by normalizing the luminescent signal from compound-treated cells to that from DMSO-treated cells. The drug concentration is expressed using a log scale. Final DMSO concentration was 0.2% in a – d . The half-maximal inhibitory concentration (IC 50 ) is indicated. Data are presented as the means ± S.E.M. ( n = 4 in c and n = 3 in d ). * P < 0.05, ** P < 0.01 or **** P < 0.0001 for each group relative to the parental cells/vehicle in c or siNT-treated cells in d . Statistical significance was calculated by one-way analysis of variance with Tukey’s multiple-comparison test
Article Snippet:
Techniques: Incubation, Liquid Chromatography with Mass Spectroscopy, Generated, Concentration Assay, Over Expression, Expressing, Glo Assay