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Figure 2 | Acta Neuropathologica Communications

Figure 2

From: Promoter DNA methylation regulates progranulin expression and is altered in FTLD

Figure 2

The DNMT-inhibitor 5-aza-2′-deoxycytidine (DAC) reduces DNA methylation in the GRN promoter and increases GRN expression in LCLs. LCL #3 and #14 were treated daily with 0.5 μM DAC for 9 days. (a) Quantitative DNA methylation analysis of amplicon A-DAC (compare Figure 1b) shows progressive demethylation in the GRN promoter region. Average DNA methylation across all 15 CpG units in the A-DAC amplicon is shown. Mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001, Student’s t-test. (b) GRN mRNA expression levels were analyzed by qPCR and normalized to PGK1. GRN mRNA expression strongly increased, while the expression of GAPDH did not change. Mean ± SD, n = 3, **p < 0.01, ***p < 0.001, ANOVA with Tukey’s Multiple Comparison test. (c) GRN secretion was quantified by ELISA and normalized to absolute cell number. Chronic treatment of LCL #3 and LCL #14 with 0.5 μM DAC for 9 days significantly increased GRN protein secretion into the cell culture supernatant, while the secretion in mock treated control cells remained low. Mean ± SD, n = 3, **p < 0.01, ***p < 0.001, ANOVA with Tukey’s Multiple Comparison test.

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