(e). n = 3, mean 6 S.D. Indoxyl sulfate-treated cells were reduced in number at 72 h compared to DMSO handle (*, P,0.05). Indoxyl sulfate-treated cell numbers have been also lowered at 72 h compared to the 8 h (a, P,0.05) and 24 h (b, P,0.05) time points. doi:ten.1371/journal.pone.0108448.gor age-dependent differences in AhR activation. Since AhR is a ligand-binding transcription factor [17], nuclear localization of podocyte AhR likely indicates the constitutive binding of endogenous ligands in adult kidneys. AhR endogenous ligands are believed to consist of eicosanoids and bilirubin [18], and indoxyl sulfate was lately reported to be an AhR ligand in liver cell lines [16]. Additional, AhR in podocytes was observed within the nucleus in vivo but within the cytoplasm in vitro. Our data suggest that podocytes may well also be a target of circulating endogenous AhR ligands.Interestingly, employing a monoclonal indoxyl sulfate antibody [43], indoxyl sulfate has been detected in podocytes as well as tubular cells in uremic rats [15]. Consequently, we propose that increased plasma levels of endogenous ligands, like indoxyl sulfate, activate podocyte AhR, thereby advertising a transcriptional system that drives progressive glomerular harm. Enhanced serum indoxyl sulfate has been linked with tubulointerstitial damage [12,13,44] and indoxyl sulfate also exacerbates glomerular injury [11]. AST-120 therapy in rodentPLOS One | plosone.orgPodocyte Injury by Indoxyl SulfateTable 1. Altered expression of podocyte function-associated genes in indoxyl sulfate-exposed human podocytes.RankGene symbolGene nameRefseqP-valueFold changePodocyte functions 1 2 three 4 5 six 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 MAP1LC3B PDPN MAP1LC3B2 WT1 FAT2 CD2AP KIRREL DES PTPRO MAP1LC3C LMX1B TJP1 MYOC NPHS1 NPHS2 MAP1LC3A EZR CD80 SYNPO2 PODXL CDH3 FAT1 PODXL2 DAG1 VIM MME CDH13 ACTN4 SYNPO microtubule-associated protein 1 light chain three beta podoplanin microtubule-associated protein 1 light chain three beta 2 Wilms tumor 1 FAT tumor suppressor homolog 2 (Drosophila) CD2-associated protein kin of IRRE-like (Drosophila) desmin protein tyrosine phosphatase, receptor form, O microtubule-associated protein 1 light chain three gamma LIM homeobox transcription aspect 1, beta tight junction protein 1 (zona occludens 1) myocilin, trabecular meshwork inducible glucocorticoid response nephrosis 1, congenital, Finnish sort (nephrin) nephrosis two, idiopathic, steroid-resistant (podocin) microtubule-associated protein 1 light chain 3 alpha ezrin CD80 molecule synaptopodin 2 podocalyxin-like cadherin 3, form 1, P-cadherin (placental) FAT tumor suppressor homolog 1 (Drosophila) podocalyxin-like 2 dystroglycan 1 (dystrophin-associated glycoprotein 1) vimentin membrane metallo-endopeptidase cadherin 13, H-cadherin (heart) actinin, alpha 4 synaptopodin NM_022818 NM_006474 NM_001085481 NM_024424 NM_001447 NM_012120 NM_018240 NM_001927 NM_030667 NM_001004343 NM_002316 NM_003257 NM_000261 NM_004646 NM_014625 NM_032514 NM_003379 NM_005191 NM_133477 NM_001018111 NM_001793 NM_005245 NM_015720 NM_001165928 NM_003380 NM_007288 NM_001257 NM_004924 NM_007286 0.1,3,5-Tribromo-2,4,6-trimethylbenzene Formula 001 0.4,6-Dimethyl-1H-indole Chemical name 003 0.PMID:24202965 000 0.111 0.359 0.070 0.159 0.568 0.514 0.370 0.487 0.177 0.755 0.274 0.181 0.514 0.068 0.377 0.484 0.215 0.040 0.007 0.214 0.034 0.000 0.001 0.000 0.000 0.000 * * * * * * * * * * * 1.36 1.28 1.22 1.20 1.13 1.09 1.08 1.07 1.04 1.03 0.98 0.97 0.96 0.93 0.93 0.92 0.92 0.91 0.91 0.90 0.88 0.87 0.87 0.85 0.85 0.83 0.72 0.67 0.Collagens 1 two three four.