FIG 4.
Krox20 is dispensable for adipogenesis in culture. (A, B) Krox20 knockout (KO) reduces growth of primary white preadipocytes. Krox20f/f primary white preadipocytes were infected with Ad-GFP or Ad-Cre. (A) Cell morphology of Krox20 KO cells under the microscope. (B) Cell growth rates: 1× 105 cells were plated, and the cumulative cell numbers were determined every day for 5 days. (C, D) Ad-GFP or Ad-Cre infected Krox20f/f primary white preadipocytes were cultured until confluence. Two days later, cells were treated with the adipogenic cocktail to induce adipogenesis. (C) Oil Red O staining at D10. (D) qRT-PCR analysis of gene expression. (E to G) Krox20f/f immortalized brown preadipocytes were infected with MSCVhygro expressing Cre. (E) Cell growth rates. (F) Oil Red O staining at day 7 of adipogenesis. (G) qRT-PCR analysis of Krox20 and adipogenesis markers at indicated time points of adipogenesis. (H, I) Krox20f/f immortalized brown preadipocytes were infected with Ad-GFP or Ad-Cre. Two days later, cells were replated, followed by adipogenesis assays. (H) Oil Red O staining at day 7 of adipogenesis. (I) qRT-PCR analysis of gene expression.