Fig. 1: Analysis of the phylogenetic tree, plasma membrane expression of SLC22A10, and uptake of organic anion substrates of the human SLC22 family.

A Multiple sequence alignments were performed with reference amino acid sequences for each anion transporter from humans and rodents, using the Clustal Omega Multiple Sequence Alignment program (https://www.ebi.ac.uk/Tools/msa/clustalo/). The dendrogram was generated from the output of the Clustal Omega alignment. Refer to the Source Data file to access the amino acid sequences for each transporter in this tree. * Human SLC22A10. B Localization of human SLC22A10 conjugated to green fluorescent protein (GFP) was examined in HEK293 cells using high-content imaging and cellular staining with the plasma membrane marker wheat germ agglutinin (WGA). Blue: DNA stain Hoechst marks the cell nucleas; Red: Plasma membrane marker WGA; Green: SLC22A10. Yellow: Merge. The results showed no colocalization of GFP-tagged SLC22A10 with WGA. The figure shows a representative image from two technical replicates. Scale bar: 10 µM. C Uptake of various radiolabeled organic anions, which are typical substrates of organic anion transporters in the SLC22A family, was assessed. Uptake was performed 48 hours after transient transfection of plasmids encoding human SLC22A10, GFP expression vector, and one other member in the SLC22A family as a positive control. Accumulation of substrates inside cells was determined after 15 minutes. The scatter plot with bars shows the fold uptake of the substrate relative to the negative control. HEK293 Flp-In cells transiently transfected with the GFP vector served as the negative control. The plot displays the mean +/− standard deviation of three or four technical replicates (n = 1 shown as a representative experiment). Source data are provided as a Source Data file. Statistical significance was determined using a one-way analysis of variance (ANOVA) with Dunnett’s multiple comparison test. n.s.: non-significant, **p value < 0.01, ****p value < 0.001. Similar results were obtained in two independent experiments.