Figure 5. Ectopic Expression of Maf1 Induces Adipogenesis in Maf1^−/− MEFs.

Maf1^−/− MEFs were infected with dox-inducible rtTA construct as control (rtTA) or co-infected with both rtTA and Maf1-HA dox-inducible constructs. Control and Maf1-HA expressed Maf1^−/− MEFs were terminally differentiated into adipocytes using a standard protocol (Figure S1C).

(A) qRT-PCR analysis of RNA expression of tRNA^Leu, tRNA_i^Met, PPARγ, C/EBPα, and FABP4 during the differentiation of either control or Maf1-HA cells that were all treated with 50 ng/mL dox from day 0 (D0) to day 12 (D12). Transcript amounts were normalized to β-actin, and the fold change was calculated relative to the amount of transcript at D0 rtTA Maf1^−/− MEF cells. Data are mean ± SD of n = 3 independent experiments. *p < 0.05, unpaired Student’s t test.

(B) Immunoblot analysis of HA-tagged Maf1, PPARγ, C/EBPα, FABP4, perilipin, and β-actin in control and Maf1 expressed Maf1^−/− MEF cells. Quantification of the changes in protein expression from the immunoblots is shown (right). Protein amounts were normalized to α-tubulin, and the fold change was calculated relative to the amount of protein in D0 rtTA control Maf1^−/− MEF cells.

(C) Oil red O staining of adipocytes that are differentiated from control and Maf1-HA expressed Maf1^−/− MEFs (left). Scale bar denotes 200 μm. Quantification of staining (right); data are mean ± SD of n = 3 independent experiments. *p < 0.05, unpaired Student’s t test.