Figure 6. Upregulation and complex formation of Fam20A and Fam20C in the lactating mammary gland.
(A) mRNA levels of Fam20C family members in the mouse mammary gland. The mammary glands from two non-lactating (Virgin) and two 10-day lactating (Lact) mice were isolated and the mRNA levels of Fam20 family members were determined by quantitative (q)RT-PCR. Data are represented as mean ± SD. (B) mRNA levels of Fam20A and Fam20C in the mammary glands of pregnant/lactating female mice at different stages. E−13, 16 or 18: embryonic day 13, 16 or 18; Lact-1, 3 or 7: 1, 3 or 7 days after lactation; Inv-1 or 7: 1 or 7 days after involution. The gestation period of mouse is 18–21 days. mRNA levels of Fam20A and Fam20C were determined by qRT-PCR. Data are represented as mean ± SD. (C) Protein levels of Fam20A and Fam20C in the mouse mammary glands. Endogenous Fam20A or Fam20C were detected from whole mammary gland lysate or the Golgi/ER-enriched fractions by western blot. (D) Co-immunoprecipitation of endogenous Fam20A and Fam20C from Golgi/ER-enriched fractions of the mouse lactating mammary gland. Polyclonal rabbit anti-Fam20C antibody was used for immunoprecipitating endogenous Fam20C. Normal rabbit IgG was used as control for immunoprecipitation.