Fig. 3.
Expression of vaspin in 293T cells, adipocytes, and stromal vascular cells. (A) Western blot analysis of mouse vaspin by using supernatants of 293T cultured cells transfected with AxCAmOL64 and polyclonal antivaspin Ab. A prominent band of ≈45 kDa is observed. (B) Western blots of purified recombinant human and mouse vaspins, derived from E. coli by using pET expression system. (C) Western blot analyses of vaspin by using sera of OLETF and LETO rats, and OLETF rats with EXE, OLETF rats administered TZD and insulin (INS). A major ≈45-kDa band and a minor ≈50-kDa band are seen at 30 wk. Band intensity is notably less at 50 wk in OLTEF rats, but it seems to be normalized with insulin and TZD treatments. (D) Expression of vaspin in mature adipocytes and stromal vascular cells isolated from epidydimal (EPI), retroperitoneal (RET), MES, and SUB WATs as analyzed by Northern and Western blot analyses. Expression is confined to adipocytes of visceral WATs and not to stromal endothelial or vascular cells. (E and F) Expression in the adipocytes was confirmed by immunofluorescence microscopy in 30-wk-old OLETF rats. (Scale bars: E, 100 μm; F, 25 μm.)