Figure 5.

WNT5A protein is retained in the ER and forms HMW aggregates in SEL1L-deficient hepatocytes

(A and B) Western blot protein analysis of total tissue lysate (A), and EndoH/PNGaseF digestion (B) of 2-month-old mouse livers (n = 5 per group, 2 independent repeats).

(C) EndoH digestion analysis of total liver lysates from 6-week-old Sel1L^f/f mice 2 weeks after tail vein injection with either AAV8-Cre or control AAV8-GFP (n = 3 per group).

(D) Western blot analysis of detergent (NP40) fractionation (S, soluble; P, pellet) of total liver lysates from 9-week-old mice (n = 3 per group). Black and red arrows mark WNT5A mobility shift. H2A and HSP90, loading controls for soluble (NP40S) and pellet (NP40P) fractions.

(E–H) Reducing and non-reducing Western blot analyses of sucrose-gradient fractionated liver lysates (E) and quantitation (F); EndoH digestion analysis of fractions #7 (G) and #13 (H) (n = 3 per group, 2 independent repeats). r and s, EndoH-resistant and -sensitive.