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. 2022 Jun 15;23(8):e52280. doi: 10.15252/embr.202052280

Figure EV3. PPARα regulates TRF transcriptional activity by binding to a PPRE in promoter of TRF.

Figure EV3

  1. Hepatic DMT1, FPN1, HEPC1 and TRF mRNA levels were measured in the indicated mice; n = 3 mice/group.
  2. Schematic of the TRF‐promoter‐PPRE.
  3. A 2,000‐base pair fragment of the promoter of mouse TRF was inserted into the pGL3 promoter vector to generate the pGL3 promoter constructs. These reporter constructs were transfected into Hep1‐6 cells. The indicated PPARα ligands were added to cell cultures 24 h before the reporter gene assay. Data were calculated as the fold induction with respect to the empty vector (pGL3 promoter luciferase vector). n = 3 biological replicates.
  4. Chromatin immunoprecipitation assays were performed on soluble formaldehyde–crosslinked chromatin isolated from untreated and GW7647‐treated WT or PPARα−/− livers with polyclonal anti‐PPARα antibodies (anti‐PPARα) or control IgG. The final DNA extraction was polymerase chain reaction‐amplified with a primer pair that covered the sequence in the promoter of Gpx4.

Data information: mRNA levels were normalized to 36B4 and are expressed relative to the mean value of the WT group; Data are presented as means ± SD. *P < 0.05, determined by ANOVA.