Figure 2.

Identification of functional ZNF750-interacting proteins. (A) Silver stain gel of Flag-HIS ZNF750 or tag-only control (CTL) tandem affinity-purified from differentiated keratinocytes. (B) Network of ZNF750-interacting proteins identified by mass spectrometry, grouped by functional category. Red lines indicate novel ZNF750 interactions, and black lines indicate published interactions. (C) Table of spectral counts and SAINT scores for 13 ZNF750-interacting proteins tested for phenotypic overlap with ZNF750 depletion. (D) Heat map of gene expression changes with depletion of ZNF750-interacting proteins by siRNA in keratinocytes induced to differentiate for 3 d with calcium, shown as number of genes with >1.5-fold expression change by qPCR in the same direction as ZNF750 depletion. Proteins above the dotted line were considered for further analysis. (E) Heat map of gene expression changes with depletion of ZNF750, RCOR1, CTBP1/2, KDM1A, or KLF4 in differentiated keratinocytes by qPCR. (F) Effects of depletion of ZNF750-interacting proteins on differentiation proteins loricrin (LOR; red) and filaggrin (FLG; green) and proliferation (Ki67; red) in regenerated organotypic epidermal tissue. Epidermal basement membrane is noted with dashed lines, and white arrowheads denote suprabasal Ki67-positive cells. Bar, 25μm. See also Supplemental Figure S2.