Fasting-Induced Hepatic Production of DHEA Is Regulated by PGC-1{alpha}, ERR{alpha}, and HNF4{alpha} -- Grasfeder et al. 23 (8): 1171 Data Supplement - Supplemental Data -- Molecular Endocrinology

Fasting-Induced Hepatic Production of DHEA Is Regulated by PGC-1 ${alpha}$ , ERR ${alpha}$ , and HNF4 ${alpha}$
Mol Endocrinol Grasfeder et al. 23: 1171

Supplemental Data

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Supplemental Figure 1 - Induction of CYP11A1 and CYP17A1 in other cell types. Real time PCR was used to measure mRNA expression in A. Primary Rat hepatocytes B. H295R cells C. AGS stomach carcinoma D. U251 glioma. Gene expression was measured in triplicate, normalized to 36B4 expression and expressed as fold induction over β-gal. Error bars represent the SEM of 3 replicates.
Supplemental Figure 2 - Induction of subsequent steroidogenic enzymes in HepG2 cells. Real-time PCR expression was used to measure mRNA expression of CYP21, cytochrome p450 family 21; 3βHSD1, 3-beta hydroxysteroid dehydrogenase type 1; 3βHSD2, 3-beta3-hydroxysteroid dehydrogenase type 2. CYP11B1, Cyptochrome p450 family 11B1. Gene expression was measured in triplicate, normalized to 36B4 expression and reported as fold induction compared to β-gal. Error bars represent the SEM of 3 replicates.
Supplemental Figure 3 - Knockdown of HNF4a target genes by siRNA in HepG2 cells. Real-time PCR expression was used to measure mRNA expression of HNF4a target genes CYP7A, cytochrome p450 family 7A, APOA4, apolipoprotein A-IV, and APOC3, apolipoprotein C-III. Error bars represent the SEM of 3 replicates.
Supplemental Figure 4 - Chromatin Immunoprecipitation of ERRalpha at sites scanned around CYP11A1. Numbering represents location of the putative ERREs relative to the start of the gene. Two additional putative ERREs were located at 19487 and 23948, and were within 130bp of the region amplified by the primer sets used for the elements at 19455 and 24042 respectively. Error bars represent the SEM of 3 replicates.
Supplemental Figure 5 - Chromatin Immunoprecipitation of ERRalpha at sites scanned around CYP17A1. Numbering represents location of the putative ERREs relative to the start of the gene. Two additional putative ERREs were located at 2970 and 12805, and were within 500bp of the region amplified by the primer sets covering the elements at 3292 and 12177 respectively. Error bars represent the SEM of 3 replicates.
Supplemental Figure 6 - Chromatin Immunoprecipitation of HNF4alpha binding to DR-1 sites scanned around CYP17A1. Numbering represents location of the putative DR-1 sites relative to the start of the gene. Error bars represent the SEM of 3 replicates.
Supplemental Table 1 - List of Primers