Journal: Advanced Science

Article Title: Selenoprotein H Functions as a PPARα Coactivator to Link Selenium Homeostasis to Hepatic Lipid Metabolism and Protect against Steatohepatitis

doi: 10.1002/advs.202519563

Figure Lengend Snippet: SELENOH binds to and activates fatty acid oxidation gene transcription in a PPARα‐dependent manner. (A) Western blot analysis of liver cytoplasmic and nuclear proteins. (B) Distribution of SELENOH signals in different genomic locations relative to known genes. (C) Barplot showing GO enrichment analysis of binding sites occupied by SELENOH. (D) Motif analysis of SELENOH‐bound sequences in mouse liver. (E) Venn diagram representing the overlapped SELENOH and PPARα peaks detected by CUT&Tag in mouse livers. (F) CUT&Tag tracks of SELENOH and PPARα peaks at the Cpt1a and Acox1 loci. (G) Luciferase reporter experiments using the PPAR response element (PPRE) in HEK293 cells. n = 3 independent culture experiments. (H) Heatmap showing ATAC‐seq signals at PPARα‐activated FAO genes in MASH control and MASH SELENOH‐OE livers. Genes shown in rows were sorted in decreasing order by signal intensity in each condition. (I and J) ATAC‐seq and RNA‐seq tracks of Cpt1a and Acox1 loci. (K) Relative mRNA levels of indicated FAO genes in liver tissues from mice injected with AAV‐TBG‐control or AAV‐TBG‐ Selenoh ‐sgRNA. n = 6 per group. (L) GO enrichment analysis of downregulated genes in SELENOH‐depleted livers. (M) Hepatic TG level of animals as indicated in (K). n = 5 per group. (N) Schematic of the experimental design: GW6471 treatment on SELENOH overexpression mice. (O) Blood AST and ALT levels, and hepatic TG level of animals as indicated. n = 5 per group. (P) Representative H&E staining of liver sections from animals as indicated. (Q) FAO activities of liver tissues from animals as indicated. n = 6 per group. (R) Relative mRNA levels of indicated FAO genes in liver tissues. n = 6 per group. (S) CUT&Tag signals of liver SELENOH at PPARα‐activated FAO genes. CUT&Tag tracks of SELENOH at Cpt1a and Acox1 loci.Values are mean ± SEM. The one‐way ANOVA with post hoc Bonferroni multiple‐comparison test (G, K, M, O, Q, and R) was used for statistical analysis.

Article Snippet: The biotinylated Acox1 ‐p probes were immobilized on Streptavidin Magnetic Beads (HY‐K0208, MCE) in DNA binding buffer (5 mmol/L Tris pH 7.5, 0.5 mmol/L EDTA, 1 mol/L NaCl).

Techniques: Western Blot, Binding Assay, Luciferase, Control, RNA Sequencing, Injection, Over Expression, Staining, Comparison

Journal: Advanced Science

Article Title: Selenoprotein H Functions as a PPARα Coactivator to Link Selenium Homeostasis to Hepatic Lipid Metabolism and Protect against Steatohepatitis

doi: 10.1002/advs.202519563

Figure Lengend Snippet: SELENOH interacts with activated PPARα and facilitates the genomic recruitment of PPARα‐P300 complex. (A) Representative immunofluorescence of SELENOH and PPARα in U2OS, Scale bars, 5 µm. (B) Endogenous co‐immunoprecipitation (co‐IP) analysis between PPARα and SELENOH in nuclear proteins from mouse livers under fasting or fed conditions. (C) Co‐IP of HA‐PPARα and FLAG‐SELENOH in HEK293T cells treated with vehicle or wy‐14643. (D) Co‐IP of different PPARα truncations with FLAG‐SELENOH in HEK293T cells treated with wy‐14643. (E) Endogenous co‐IP of PPARα and P300 in liver nuclear proteins from MASH control and MASH SELENOH‐OE mice. (F) Heatmap showing CUT&Tag signals of P300 at PPARα‐activated FAO genes in MASH control and MASH SELENOH‐OE livers. (G) Endogenous co‐IP of PPARα and P300 in liver nuclear proteins from mice injected with AAV‐TBG‐control or AAV‐TBG‐ Selenoh ‐sgRNA under the fasting condition. (H) Heatmap showing CUT&Tag signals of P300 at PPARα‐activated FAO genes in livers from mice injected with AAV‐TBG‐control or AAV‐TBG‐ Selenoh ‐sgRNA under the fasting condition. (I) Schematic of DNA pulldown assay. (J) Western blot analysis of biotin‐ Acox1 promoter ( Acox1 ‐p) pull‐down of nuclear protein extracts from indicated liver tissues. (K) Heatmap showing CUT&Tag signals of PPARα in MASH control and MASH SELENOH‐OE livers. (L) CUT&Tag tracks of P300 and PPARα peaks at the Cpt1a locus. (M) Heatmap showing CUT&Tag signals of PPARα in livers from mice injected with AAV‐TBG‐control or AAV‐TBG‐ Selenoh ‐sgRNA under the fasting condition. (N) CUT&Tag tracks of P300 and PPARα peaks at the Cpt1a locus. (O) Co‐IP of HA‐PPARα and FLAG‐SELENOH SXXS in HEK293T cells treated with vehicle or wy‐14643. (P) Relative mRNA levels of indicated FAO genes in primary hepatocytes from mice injected with AAV‐TBG‐control, AAV‐TBG‐SELENOH, or AAV‐TBG‐SELENOH SXXS . n = 6. (Q) Luciferase reporter experiments using the PPAR response element (PPRE) in HEK293T cells. Cells were transfected with indicated plasmid with or without 1 µM selenium (Se) treatment. n = 3 independent culture experiments.Values are mean ± SEM. The one‐way ANOVA with post hoc Bonferroni multiple‐comparison test (P and Q) was used for statistical analysis.

Article Snippet: The biotinylated Acox1 ‐p probes were immobilized on Streptavidin Magnetic Beads (HY‐K0208, MCE) in DNA binding buffer (5 mmol/L Tris pH 7.5, 0.5 mmol/L EDTA, 1 mol/L NaCl).

Techniques: Immunofluorescence, Immunoprecipitation, Co-Immunoprecipitation Assay, Control, Injection, Western Blot, Luciferase, Transfection, Plasmid Preparation, Comparison