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. Author manuscript; available in PMC: 2022 Feb 1.
Published in final edited form as: Arterioscler Thromb Vasc Biol. 2020 Dec 10;41(2):783–795. doi: 10.1161/ATVBAHA.120.315310

Figure 4. Transcription factor EB (TFEB) upregulates insulin receptor substrate 2 (IRS2) expression at the transcriptional level.

Figure 4.

A, Chromatin immunoprecipitation sequencing (ChIP-seq) data were obtained from the Gene Expression Omnibus (GEO, accession number GSE88896) and were analyzed for TFEB binding peaks in the promoter of human IRS2 gene. B, Human coronary artery endothelial cells (HCAECs) were infected with Ad-LacZ (20 MOI) or Ad-Flag TFEB (20 MOI) for 48 hours. The binding of TFEB to the IRS2 promoter was measured by ChIP assays using the antibody against Flag (n=3/each group). C, Hela cells were used as a tool for the reporter gene assay. The cells were transfected with pGL4.10 Luciferase reporters driven by wild type (WT, 500 bp IRS2 promoter region: −440bp~+59bp) or deletion (del, −65 bp~−59 bp deletion corresponding to the TFEB binding site) form of IRS2 promoter for 6 hours, and then the cells were infected with Ad-LacZ (10 MOI) and Ad-TFEB (10 MOI). Forty-eight hours later, luciferase activity was measured and normalized by Renilla activity (n=8/each group). Data in B and C are presented as mean ± SEM; **P <0.01 using 2-way ANOVA followed by Bonferroni test.