Fig. 8.

Localization of LRP2 in primary chicken kidney epithelial cells and induction by estrogen. The cells were isolated and prepared for immunocytochemistry with anti-chicken LRP2 antiserum or preimmune serum (Pre-S) as described in Material and methods. A: Immunofluorescence (magnification, 40 ×). B: Quantitative real-time PCR of LRP2 mRNA in primary chicken kidney epithelial cells. Cells were treated with 50 nM estrogen and harvested after 24 h. All values are expressed as means ± SEM of mRNA levels relative to those of β-actin; experiments were performed in triplicate. Data were analyzed by Student's t-test (*, p < 0.05). C: Determination of LRP2 protein levels in lysates of primary chicken kidney epithelial cells treated (lanes 2 and 3) or not treated for 24 h (lanes 1 and 2) with 50 nM estrogen. 50 μg of protein/lane was separated by 6% (or 12%, respectively) SDS-PAGE under non-reducing conditions and processed for Western blot analysis with rabbit anti-ggLRP2 antiserum (1:1000). The anti-panGAPDH antibody (1:20,000) was used to detect the product of the housekeeping gene.