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. 2017 Dec 12;10:413. doi: 10.3389/fnmol.2017.00413

Figure 6.

Figure 6

Epigenetic silencing of the Creb3l1 gene by CpG promoter methylation. (A) Expression of Creb3l1 and Nr4a1 in different mammalian cell lines presented as CT value. (B) Time course of FSK treatment in three different cell lines. (C) The effect of Nr4a1 knockdown on Creb3l1 expression in Mcf-7 and N2a cells. (D) Identification of a conserved CpG island in the human and rodent Creb3l1 promoters (4.9 kb). (E) Methylation status of the Creb3l1 promoter CpG island in three different cell lines. (F) Luciferase assays of methylated Creb3l1 luciferase reporter constructs with overexpression of Nr4a1 or 4 h FSK treatment. (G) qRT-PCR analysis of Creb3l1 and Nr4a1 in cell lines treated with doses of 2.5–10 μM of 5-Aza-2′deoxycytidine for 72 h. (H) FSK stimulation (4 h) of cells treated with 5-Aza-2′deoxycytidine (72 h, 5 μM). (I) The effect of 5-Aza-2′deoxycytidine and FSK treatment on expression of arginine vasopressin (Avp) in N2a cells. Increase of Avp mRNA expression by overexpression of a constitutively active (CA) mutant of Creb3l1 in N2a cells. TSS, transcription start site. Values are means + SEM of n = 3−4 per group. *p < 0.05, **p < 0.01, ***p < 0.001.