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. Author manuscript; available in PMC: 2016 Jul 1.

Published in final edited form as: Gastroenterology. 2015 Mar 30;149(1):151–162. doi: 10.1053/j.gastro.2015.03.046

(a) Genotyping and RT-PCR of Atg7 in mouse intestinal epithelial cells (Atg7^−/−). (b) Western blot and quantification of p62, LC3I, and LC3III in Atg7^−/− mouse intestine treated with vehicle or rapamycin. (c) Western blot analysis of NHE3 and other proteins as indicated in control and Atg7^−/− mice with and without rapamycin treatment, and surface biotinylation assay to evaluate the surface levels of NHE3 and NHERF1 in the membrane fraction. (d) Quantification of NHE3 and other proteins in control (open bar) and Atg7^−/− mice (filled bar) are shown, as is quantification of NHE3 and NHERF1 in the membrane fraction. The comparisons that show statistically significant differences among the groups, with p < 0.05, are indicated as follows: *:Atg7^−/−/Veh versus Atg7^F/F/veh or Atg7^−/−/Rapa versus Atg7^F/F/Rapa; **: Atg7^F/F/veh versus Atg7^F/F/Rapa; and #: Atg7^−/−/Rapa versus Atg7^−/−/veh (N=3, one-way ANOVA with Bonferroni’s multiple comparison test). (e–f) Representative images (e) and quantification (f) of immunostaining of NHE3 in the ileum from Atg7^F/F and Atg7^−/− mice (*p<0.05, t-test, N= 3).

(a) Genotyping and RT-PCR of Atg7 in mouse intestinal epithelial cells (Atg7^−/−). (b) Western blot and quantification of p62, LC3I, and LC3III in Atg7^−/− mouse intestine treated with vehicle or rapamycin. (c) Western blot analysis of NHE3 and other proteins as indicated in control and Atg7^−/− mice with and without rapamycin treatment, and surface biotinylation assay to evaluate the surface levels of NHE3 and NHERF1 in the membrane fraction. (d) Quantification of NHE3 and other proteins in control (open bar) and Atg7^−/− mice (filled bar) are shown, as is quantification of NHE3 and NHERF1 in the membrane fraction. The comparisons that show statistically significant differences among the groups, with p < 0.05, are indicated as follows: *:Atg7^−/−/Veh versus Atg7^F/F/veh or Atg7^−/−/Rapa versus Atg7^F/F/Rapa; **: Atg7^F/F/veh versus Atg7^F/F/Rapa; and #: Atg7^−/−/Rapa versus Atg7^−/−/veh (N=3, one-way ANOVA with Bonferroni’s multiple comparison test). (e–f) Representative images (e) and quantification (f) of immunostaining of NHE3 in the ileum from Atg7^F/F and Atg7^−/− mice (*p<0.05, t-test, N= 3).