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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 2. — Genetic disruption of the ZFP423–EBF2 interaction in adipocytes leads to thermogenic WAT remodeling. (A) Components of the Adipo-Zfp423^WT-Avi and Adipo-Zfp423^H1285N-Avi mouse models. Quadruple transgenic mice consist of Adiponectin^rtTA and TRE-Cre transgenes, one floxed Zfp423 allele (Zfp423^lopx/+), and one allele engineered to express either ZFP423^WT-Avi (Zfp423^WT-Avi/+) or ZFP423^H1285N-Avi (Zfp423^H1285N-Avi/+). Dox-chow diet feeding results in the inactivation of one Zfp423 allele in adiponectin-expressing mature adipocytes, with the other allele expressing either ZFP423^WT-Avi in Adipo-Zfp423^WT-Avi mice or ZFP423^H1285N-Avi in Adipo-Zfp423^H1285N-Avi mice. (B–D) Representative bright-field images of H&E-stained sections of iWAT depots obtained from adult mice with the indicated genotypes after 4 wk of Dox-chow diet feeding at room temperature. Scale bar, 200 μm. Representative photographs of dissected whole iWAT depots are shown. (E) Relative mRNA levels of thermogenic genes in iWAT from adult mice with the indicated genotypes after 4 wk of Dox-chow diet feeding at room temperature. Bars represent mean ± SEM. (*) P < 0.05 for Adiponectin^rtTA; Zfp423^loxp/loxP versus Adiponectin^rtTA; TRE-Cre; Zfp423^loxP/loxP by one-way ANOVA. n = 5 biological replicates. (F–I) Representative bright-field images of H&E-stained sections of iWAT depots obtained from adult mice with the indicated genotypes after 4 wk of DOX-chow diet feeding at room temperature. Scale bar, 200 μm. Representative photographs of dissected whole iWAT depots are shown. (J) Relative mRNA levels of thermogenic genes in iWAT from adult mice with the indicated genotypes after 4 wk of Dox-chow diet feeding at room temperature. Bars represent mean ± SEM. (*) P < 0.05 for Adiponectin^rtTA; Zfp423^loxp/WT-Avi versus Adiponectin^rtTA; TRE-Cre; Zfp423^{loxP/H1285N-Avi} by one-way ANOVA. n = 4 biological replicates. (K) Western blot analysis of UPC1 protein levels in whole iWAT depots isolated from adult mice with the indicated genotypes after 4 wk of Dox-chow diet feeding at room temperature.