Figure 6.

CRT downregulation promotes lipid accumulation in mice primary hepatocytes and its levels are inhibited in the db/db mice liver (A) Mice primary hepatocytes were transfected with either the scramble or CRT siRNA or p53 siRNA and stained with Bodipy and DAPI. Bodipy fluorescence was quantified and the data are given below. (B) 30 µg protein from normal and db/db mice (n = 3 ) liver were run on SDS-PAGE, transferred to nitrocellulose membranes and probed with antibodies against SREBP-1c, CRT and p53. Vinculin was used as the loading control and densitometric analyses of the same are given below. Experiment from each animal was run thrice and a representative blot is shown. (C) Schema demonstrating that CRT inhibition leads to p53 downregulation, that due to its decreased occupancy of the SREBP-1c promoter, facilitates its upregulation and subsequently promotes lipid synthesis and accumulation. All values are means±SEM from 3 independent experiments. **p < 0.01 and *p < 0.05 as compared to scramble transfected cells.