Figure 7. Intact caveolae are required for PTRF tyr-phosphorylation, nuclear translocation and ribosomal function.
(A) Isolated mouse primary adipocytes treated with beta-cyclodextrin (Beta-CD) or not were fractionated into crude lipid rafts and non lipid rafts fractions. Lysate from both fractions were run on SDS-PAGE and blotted with indicated antibodies. (B) Nuclear fraction lysates from beta-CD (45-min) and insulin (30-min) treated or not adipocyte were run on SDS-PAGE and blotted with indicated antibodies, histone-H3:H3. (C) Whole cell lysates from B were separated by SDS-PAGE and blotted with phosphotyrosine antibody (4G10). Arrows indicate the size of 180 and 95 kDa, which possibly corresponds to IRS-1/2 and beta-subunit of insulin receptor respectively. The whole cell lysates were also immunoprecipitated by PTRF antibody followed by SDS-PAGE and blotted with indicated antibodies. (D) 47S levels of the samples from B were measured by RT-qPCR.