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. 2021 Nov 1;35(21-22):1461–1474. doi: 10.1101/gad.348780.121

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© 2021 Shao et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 1. — Identification of zinc fingers (ZFs) mediating ZFP423 interaction with EBF2. (A) Schematic representation of 30 zinc finger (ZF) motifs within murine ZFP423. The previously identified EBF family interaction domain is highlighted in blue. ZFs 28–30 were individually mutated through histidine-to-asparagine substitutions at the indicated histidine residues. (B) Western blot analysis of the indicated proteins in whole-cell lysates (top) and ZFP423 immunoprecipitates (bottom) from differentiated immortalized brown stromal vascular cells following sequential transduction with retrovirus carrying Ebf2 and then retrovirus expressing FLAG-tagged (FL) ZFP423 variants. (C) Relative mRNA levels of thermogenic genes (top) and common adipocyte-selective genes (bottom) in cultures of in vitro differentiated immortalized brown stromal vascular cells expressing the indicated ZFP423 variants. Bars represent mean + SEM. (*) P < 0.05 by one-way ANOVA. n = 3 biological replicates.