Figure 1. Effect of in vitro acute leptin treatment on aquaglyceroporins AQP3 and AQP7 expression and subcellular localization in murine adipocytes.

Bar graphs show transcript and protein levels of AQP3 (A, C) and AQP7 (B, D) in differentiated murine adipocytes obtained from subcutaneous white adipose tissue (WAT) of wild type mice under basal conditions and after leptin (10 nmol/L) treatment for 24 h. The gene and protein expression in unstimulated cells was assumed to be 1. Representative blots are shown at the bottom of the figure. (E) Immunocytochemical detection of the AQP3 and AQP7 proteins in differentiated murine 3T3-L1 adipocytes (day 10) under basal conditions (upper panels) and after the stimulation for 4 h with leptin (10 nmol/L) (lower panels). Images were taken from the basal planes of the cells. Representative images of at least three separate experiments are shown. (F) Glycerol release from murine subcutaneous adipocytes under basal conditions (control) and after leptin (10 nmol/L)-induced stimulation without or with preincubation with HgCl₂ (0.3 mmol/L), a nonspecific AQP inhibitor, or with CuSO₄ (0.1 mmol/L), a selective AQP3 inhibitor. Differences between groups were analyzed by Student’s t test or one-way ANOVA followed by Tukey’s test. *P < 0.05 vs. control unstimulated cells; ††P < 0.01 vs. adipocytes stimulated with leptin.