(a–b) Western blot analysis (a) and quantification (b) of NHE3, NHERF1, CFTR, Ezrin, p-Ezrin, S6, p-S6, Akt, p-Aktser473 in the small intestine from vehicle (Veh)- and rapamycin (Rapa)-treated rats. (c–d) Surface biotinylation assay (c) and quantification (d) for surface expression of NHE3 and NHERF1 in the membrane fraction. (e) Representative images and quantification of NHE3 at the apical membrane by immunostaining of NHE3 in mid-villus of the ileum from vehicle- and rapamycin-treated rats. (f) Western blot and quantification of p62, LC3I and LC3II in control and rapamycin-treated rat intestine (*p<0.05, t-test, N=3).