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. Author manuscript; available in PMC: 2021 May 1.
Published in final edited form as: Hypertension. 2020 Mar 30;75(5):1233–1241. doi: 10.1161/HYPERTENSIONAHA.120.14642

Figure 3. H3K27me3 bound to Klotho promoter and downregulated Klotho gene expression.

Figure 3.

(A) Dose-dependent upregulation of sKL and downregulation of H3K27me3 by GSK343 in primary cultured mouse renal tubule cells. (B) Dose-dependent upregulation of sKL and downregulation of H3K27me3 by EED226 in primary cultured mouse renal tubule cells. (C) Klotho promoter, basal promoter activity of human Klotho gene is controlled by the DNA sequence located between Sac2 and Pml1 site of Klotho promoter region; pHK-Sac2, effect of EED226 and GSK343 on the promoter activity of pHKP-Sca2 construct; pHK-Pml1, upregulation of Klotho promoter activity drived by pHKP-Pml1 construct in the presence of PRC2 inhibitor EED226 and/or GSK343. (D) ChIP assay of H3K27me3 binding to Klotho basal promoter region and ChIP qPCR quantitation of the effect of EED226 and/or GSK343 on H3K27me3 binding to Klotho promoter in vitro. (E) ChIP assay of H3K27me3 binding to Klotho basal promoter region in vivo and ChIP qPCR quantitation of H3K27me3 binding to Klotho promoter in the kidney of WT and Klotho mutant mice in vivo. N=4. Data are expressed as the mean ± S.D. * p < 0.05, or ** p<0.01 by unpaired t-test and multiple groups by one-way analysis of variance.