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10. Additional file 2 of Klf4-Sirt3/Pparα-Lcad pathway contributes to high phosphate-induced lipid degradation

Author

Yu, Angen, Xu, Yichuang, Hogstrand, Christer, Zhao, Tao, Tan, Xiao-Ying, Wei, Xiaolei, Song, Yu-Feng, and Luo, Zhi

Abstract

Additional file 1: Supplemental Text S1. Yellow catfish primary intestinal epithelial cells (IECs) isolation and culture. Supplemental Table S1. Feed formulation and proximate analysis of experimental diets. Supplemental Table S2. Primers used for quantitative real-time PCR analysis. Supplemental Table S3. Primers used for plasmid construction of expression vector. Supplemental Table S4. Primers used for plasmid construction of si-klf4. Supplemental Table S5. Primers used for plasmid construction of promoters. Supplemental Table S6. Primers used for site-mutation analysis. Supplemental Table S7. Primers used for electrophoretic mobility-shift assay. Supplemental Table S8. Effects of dietary phosphorus supplementation on growth performance and feed utilization of yellow catfish. Supplemental Figure S1. MTT assay of primary IECs under Pi incubation. Supplemental Figure S2. Immunofluorescent staining of KLF4 protein under Pi incubation. Supplemental Figure S3. Nucleotide sequence of yellow catfish sirt3 promoter. Numbers are relative to the transcription start site (+1). Supplemental Figure S4. ERRα response elements (ERE) located at −1443 bp to −1457 bp and KLF4 response elements (KRE) located at −406 bp to −419 bp of pparα promoter. Supplemental Figure S5. Relative luciferase activity of sirt3 promoter after the incubation with different overexpression vectors. Supplemental Figure S6. The immunoprecipitation experiment for the analysis of KLF4 and ERRα binding with SIRT3. Supplemental Figure S7. Nucleotide sequence of yellow catfish pparα promoter. Numbers are relative to the transcription start site (+1). Supplemental Figure S8. KLF4 response elements (ERE) located at −1443 bp to −1457 bp (KRE1) and −406 bp to −419 bp (KRE2) of pparα promoter. Supplemental Figure S9. Nucleotide sequence of yellow catfish lcad promoter. Numbers are relative to the transcription start site (+1).

Published
2023
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30. Functional Analysis of Steroidogenic Factor 1 (sf-1) and 17α-Hydroxylase/Lyase (cyp17α) Promoters in Yellow Catfish Pelteobagrus fulvidraco.

Author

Lv, Wu-Hong, Chen, Guang-Hui, Zhuo, Mei-Qin, Xu, Yi-Huan, Xu, Yi-Chuang, and Tan, Xiao-Ying

Subjects
FLATHEAD catfish, FACTOR analysis, FUNCTIONAL analysis, LEPTIN, PEROXISOME proliferator-activated receptors, PROMOTERS (Genetics)
Abstract

The present study was performed to clone and characterize the structures and functions of steroidogenic factor 1 (sf-1) and 17α-hydroxylase/lyase (cyp17α) promoters in yellow catfish Pelteobagrus fulvidraco, a widely distributed freshwater teleost. We successfully obtained 1981 and 2034 bp sequences of sf-1 and cyp17α promoters, and predicted the putative binding sites of several transcription factors, such as Peroxisome proliferator-activated receptor alpha (PPARα), Peroxisome proliferator-activated receptor gamma (PPARγ) and Signal transducer and activator of transcription 3 (STAT3), on sf-1 and cyp17α promoter regions, respectively. Overexpression of PPARγ significantly increased the activities of sf-1 and cyp17α promoters, but overexpression of PPARα significantly decreased the promoter activities of sf-1 and cyp17α. Overexpression of STAT3 reduced the activity of the sf-1 promoter but increased the activity of the cyp17α promoter. The analysis of site-mutation and electrophoretic mobility shift assay suggested that the sf-1 promoter possessed the STAT3 binding site, but did not the PPARα or PPARγ binding sites. In contrast, only the PPARγ site, not PPARα or STAT3 sites, was functional with the cyp17α promoter. Leptin significantly increased sf-1 promoter activity, but the mutation of STAT3 and PPARγ sites decreased leptin-induced activation of sf-1 promoter. Our findings offered the novel insights into the transcriptional regulation of sf-1 and cyp17α and suggested leptin regulated sf-1 promoter activity through STAT3 site in yellow catfish. [ABSTRACT FROM AUTHOR]

Published
2021
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