Fig. 4: rs6048205-G enhanced RXRA binding to activate FOXA2 expression in pancreatic progenitor stage abnormally. | Nature Communications

Fig. 4: rs6048205-G enhanced RXRA binding to activate FOXA2 expression in pancreatic progenitor stage abnormally.

From: A noncoding variant confers pancreatic differentiation defect and contributes to diabetes susceptibility by recruiting RXRA

Fig. 4

a Firefly luciferase/Renilla luciferase results of rs6048205 alleles in PANC1, DE, PP1, PP2 cells (n = 4 independent experiments). b The sequence around rs6048205 and the consensus RXRA binding motif from JASPAR. c EMSA results show purified RXRA protein binding to biotin-labeled probes containing two allelic variants of rs6048205 (n = 3 independent experiments). d Firefly luciferase/Renilla luciferase results show RXRA overexpression regulation on rs6048205 in 293T (n = 4 independent experiments). e Allele-specific ChIP-qPCR assay for comparing RXRA binding between allele A and allele G in Huh7 cells, which is heterozygous in rs6048205 (n = 3 independent experiments). f Time-course quantification of relative mRNA levels of RXRA along with FOXA2 detected by RT-qPCR analysis (n = 3 independent experiments). g Representative immunofluorescence images of DE and PP cells stained by RXRA with SOX17 and PDX1, respectively (n = 3 independent experiments). Scale bar, 50 µm. h mRNA levels of FOXA2 (n = 4 independent experiments) and NKX6-1 (n = 3 independent experiments) in PP2 cells treated with different dosages of RXR agonists bexarotene (Bexa) and LG000268 (LG268) detected by RT-qPCR analysis. i Protein levels of FOXA2, PDX1, and NKX6-1 in PP2 cells treated with the highest dosage of RXR agonists bexarotene (Bexa, 200 nM) and LG000268 (LG268, 40 nM) detected by Western blot (n = 3 independent experiments). GAPDH was used as a loading control. j Representative immunofluorescence images of PDX1 and NKX6-1 staining in PP2 cells treated with the highest dosage of RXR agonists bexarotene (Bexa, 200 nM) and LG000268 (LG268, 40 nM). k mRNA levels of FOXA2 and NKX6-1 in G/G PP2 cells treated with different dosage of RXR inhibitor UVI3003 (UVI) detected by RT-qPCR analysis (n = 4 independent experiments). Data are presented as the mean ± SD. Statistical significance was determined using the unpaired, two-tailed t-test in (a, d, e, h, k) (ns not significant). Source data are provided as a Source Data file.

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