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. 2015 Nov 12;310(2):G64–G69. doi: 10.1152/ajpgi.00367.2015

Fig. 1.

Fig. 1.

Identification of sodium/hydrogen exchanger (NHE) 8 expression in goblet cells. A: PCR detection of NHE isoform expression in HT29-MTX cells. Total RNA was isolated from cells and used for 1st-strand cDNA synthesis. Real-time PCR was performed using human NHE2, NHE3, NHE8, and TATA-binding protein (TBP) primers. Data are presented as means ± SE from 6 separate experiments. *P < 0.05 for Caco-2 cells vs. HT29-MTX cells. B: NHE8 protein localization in goblet cells. Mouse colonic tissue section and HT29-MTX cells were reacted with NHE8 antibody following the procedure described in materials and methods. NHE8 is labeled with red, and nuclei are labeled with blue. C: characterization of functional NHE8 in HT29-MTX cells. Cells were loaded with 40 mM NH4Cl for 5 min and then washed with sodium-free HBSS before addition of sodium-containing HBSS. The rate of intracellular pH (pHi) recovery was analyzed during the initial 60 s following addition of sodium-containing HBSS in the presence or absence of NHE inhibitor HOE-694 (0, 1, and 10 μM). HOE-694 was included both during the NH4Cl washout phase and the recovery phase. Data were collected from 19 to 29 cells for each group. *P < 0.00001 for 1 μM HOE-694 vs. others; #P < 0.00001 for 10 μM HOE-694 vs. others.