Unraveling the functional role of the orphan solute carrier, SLC22A24 in the transport of steroid conjugates through metabolomic and genome-wide association studies
Fig 4
Inhibition studies of various anions in HEK293 Flp-In cells stably expressing SLC22A24.
(A) Inhibition of [3H]-taurocholic acid uptake (with 1 μM unlabeled taurocholic acid) in cells stably expressing SLC22A24 by various bile acids, steroids, steroid conjugates, dicarboxylic acids, non-steroidal drugs, and other substances. All compounds were screened at 100 μM, and for some compounds, also at 1 mM. Uptake of [3H]-taurocholic acid was stopped after 10 min. Values represent the mean ± S.E.M. (from at least one experiment with three replicate wells each time). *Compounds that are statistically significant in the metabolomic analysis. #Steroidal compounds that are statistically significant in the genome-wide association studies. (B) Inhibition plots for SLC22A24-mediated taurocholic acid uptake in HEK293 Flp-In cells stably expressing SLC22A24. Cells were incubated with HBSS buffer containing taurocholic acid (1 μM) for 10 min with or without various concentrations of the compounds. Values are presented as mean ± S.D. of taurocholic acid uptake from three replicate wells determined in a single experiment. The experiments were repeated once and similar results were obtained. Representative curves of the SLC22A24-mediated taurocholic acid uptake inhibition by steroid conjugates (progesterone sulfate, etiocholanolone glucuronide), bile acid, and drugs (chenodeoxycholic acid, lesinurad, rosiglitazone). Underlying data are provided in S1 Data. See Table 3 for the IC50 values.