Figure 2

Tgm2−/− MEFs show accelerated adipogenesis. (a) Immunofluorescence staining for lipid with Bodipy 493/503 (green) during differentiation of Tgm2+/+ and Tgm2−/− MEFs. Lipid accumulation is visible on day 3 in Tgm2−/− MEFs compared with Tgm2+/+ MEFs which show lipids only on day 4–5 onwards. Nuclei were visualized with DAPI (blue). Scale bar equals 50 μm. (b) mRNA expression of Pparγ and Cebpα from Tgm2+/+ and Tgm2−/− MEFs on day 3, show a significant increase in Tgm2−/− MEFs. The relative quantity of mRNA expression was normalized to 18 S. Error bars±SD (n=3), **P<0.01. (c) Nuclear translocation of PPARγ and C/EBPα (colocalization with DAPI in pink) in Tgm2+/+ and Tgm2−/− MEFs on day 3 show increased activation of transcription factors. Nuclei stained with DAPI (blue). Scale bar equals 50 μm. (d) Quantification of PPARγ and C/EBPα positive nucleus per total number of cells in Tgm2+/+ and Tgm2−/− MEFs on day 3 shows a dramatic and significant increase in Tgm2−/− MEFs. Error bars±SEM (n=3), *P<0.05. (e) Western blot analysis of PPARγ and C/EBPα in total cell lysates on days 3 and 4 show increased PPARγ and C/EBPα protein levels in Tgm2−/− MEFs; actin used as loading control