ZNF750 regulates keratinocyte terminal differentiation.

<p>(A) Gene Ontology terms for the 256 genes that were downregulated in <i>ZNF750</i> silenced cultures vs. control (P-value<0.05, FC≥2), with Bonferroni-corrected p-value<0.001. (B) QRT-PCR mRNA expression of a selected group of terminal differentiation genes in control vs. <i>ZNF750</i> silenced cultures, that showed downregulation fold change≥2. Error bars represent mean values±SD, N = 2. (C) mRNA expression (QRT-PCR) of a selected group of terminal differentiation genes in differentiated cells vs. undifferentiated HaCaT cells that overexpress <i>ZNF750</i> or an empty control vector (CTL). Cells in undifferentiated state were maintained at sub-confluence in low calcium media. Expression levels are shown relative to those of Ca²⁺-induced differentiated HaCaT cultures at day twelve (100%). Two sets of primers targeting ZNF750 were used: “ZNF750 endogenous” PCR amplifies the ZNF750 5′ UTR, representing the ZNF750 endogenous expression only, and serves as an internal control demonstrating the cells' undifferentiated state. The ZNF750 Exon2-3 primer set PCR amplifies ZNF750 coding sequence common to the endogenous and exogenous ZNF750 molecules. Note that in the CTL control group mRNA levels of all tested markers were practically null. Error bars represent mean values±SD, N = 2. (D) Schematic representation: ZNF750 expression is gradually augmented throughout keratinocyte differentiation, inducing expression of late differentiation markers.