Western blot analysis of mouse FD in serum and adipose tissue. (A)
Untreated serum sample from WT mouse (0.3 µl) and also of recombinant
mouse FD (30 ng) were treated with PNGase F. (B) Western blot of mouse FD in
various adipose tissues: EWAT (Epididymal White Adipose Tissue), IWAT (Inguinal
White Adipose Tissue), BAT (Brown Adipose Tissue). (C) Western blot of 1)
undifferentiated MEFS prior to differentiation (UN0, lane 1), 2) MEFs
differentiated into white adipocytes with the Dexamethasone/Insulin/IBMX
protocol (DIX) for 9 d (DIX9, lane 2), 3) MEFs differentiated into brown
adipocytes with DIX plus the PPAR-γ agonist troglitazone (DIXγ)
protocol for 9 d (DIXγ9, lane 3) and 4) undifferentiated MEFs at the
conclusion of the differentiation period (UN9, lane 4). (D) Western blot of
mouse FD in various fat tissues (EWAT, IWAT, and BAT) before and after treatment
with PNGaseF. These results demonstrate that mouse FD both in serum and adipose
tissues is highly and variably N-linked glycosylated.