ER stress response in 3T3-A212P pre-adipocytes.

<p>(A) 3T3-TRE-WT and 3T3-TRE-A212P cells were treated with 100 ng/ml of Dox at the indicated time points. Total cell lysate was collected and immuno-blotting was performed by using the Myc antibody. The asterisk indicates a nonspecific band as a loading control. (B) 3T3-TRE-WT and 3T3-TRE-A212P cells were treated with 50 or 100 ng/ml Dox after cells reached 100% confluency. Two days later, total cell lysate was collected and immuno-blotting was performed using BiP and CHOP antibodies. β-actin was used as the loading control. (C) Quantification of protein expression levels in (B). Data are presented as mean ± SD from a representative of six independent experiments. Comparison was made between 3T3-TRE-WT and 3T3-TRE-A212P cells receiving the same dosage of Dox treatment. *p<0.05, **p<0.01. (D) 3T3-TRE-WT and 3T3-TRE-A212P cells were treated with 100 ng/ml Dox after cells reached 100% confluency. Two days later, expression levels of ER stress markers BiP and CHOP were measured by real-time qPCR. Data are presented as mean ± SD from three independent experiments. Comparison was made between 3T3-TRE-WT and 3T3-TRE-A212P cells receiving the same dosage of Dox treatment. *p<0.05, **p<0.01. (E) A proposed model on how Seipin-A212P inhibits adipogenesis. Seipin-A212P induction is associated with inflammatory responses in the early predifferentiation stage, which in turn inhibits PPARγ expression and suppresses adipogenesis.