QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

Help viewing high resolution images Return to article

Fig. 2. Effect of ammonium chloride and of chloroquine on rPRL cleavage in rat MECs. Electrophoresis under reducing conditions and immunoblotting analysis of rPRL forms detectable in mammary epithelial cells and in their media upon incubation with 23 kDa rPRL in the presence or absence of 10 mM ammonium chloride (NH₄Cl) or 10 µM chloroquine (ClQ) as indicated. Mammary fragments were preincubated for 30 minutes at 37°C in the presence or absence of the drug, then 5 µg/ml rPRL was added or not for 30 minutes at 20°C in order to accumulate the hormone intracellularly and further incubated for 60 minutes at 37°C. Incubation media and fragments were treated for immunoblotting as described in Materials and Methods. (A) Immunoblotting analysis of rPRL forms in homogenates from tissue fragments (cells) incubated in the presence of weak bases. The 23 kDa form of rPRL was detectable in tissue incubated in the presence of exogenous PRL. (B) Immunoblotting analysis of rPRL forms in the incubation medium of tissues fragments (medium) incubated in the presence of weak bases. In addition to the 23 kDa form of rPRL, a form with an M_r of about 18 kDa and a much more abundant form with an M_r of 16 kDa were detectable. (C) Immunoblotting analysis of rPRL forms in homogenate and medium of tissue fragments incubated in Hanks' medium in the presence of exogenous rPRL. In tissue fragments (cells) the 23 kDa was detectable. In the medium, the 23 kDa and the 16 kDa forms were detectable. Positions of the molecular mass markers (kDa) are indicated on the left.

Return to article