Figure 5

N-glycosylation of Abcg5/sterolin-1 analyses in Abcg8-/- mouse liver. Mouse liver homogenate stained with SC anti-Abcg5/sterolin-1 after treatment with Endo-H or PNGaseF shows a 75 kDa band present in both genotypes, which is resistant to deglycosylation (panel a). Staining of wild-type liver homogenate with preimmune serum showed no detectable bands. Lower portion of panel (a) shows the same aliquots stained for anti-transferrin as a control for deglycosylation. AMC anti-Abcg5/sterolin-1 staining of mouse liver homogenate shows a 'mature' ~90 kDa band in the wild-type mice but not in the Abcg8-/- mice (panel b). A 75 kDa form is present which is sensitive to deglycosylation. Mouse liver homogenate stained with UTSW anti-Abcg5/sterolin-1 shows a 'mature' ~90 kDa band and an 'immature' 75 kDa band in the wild-type mice but no signal is detected in the Abcg8-/- mice. Treatment with Endo-H or PNGaseF results in a lower molecular weight protein in the wild-type mice. Abcg5/Abcg8-/- liver homogenate used for negative control.