Skip to main content
. 2004 Dec 10;101(51):17705–17710. doi: 10.1073/pnas.0407974101

Fig. 2.

Fig. 2.

Decreased expression levels of ERM proteins in EBP50(–/–) BBM. (A) Schematic protocol of BBM preparation. The protein concentration was measured in each boxed fraction, and samples containing 30 μg of proteins [except from S4 (supernatant 4), which contained only 1.5 μg of proteins] were loaded on polyacrylamide gels. TL, total tissue lysates. P4 (pellet 4) corresponds to the BBM fraction. (B) Western blot analysis of kidney and small intestine BBM fractionation samples from EBP50 (+/+) and (–/–) mice was performed with the indicated antibodies. The quantification analysis (Right) was performed with the imagej program (National Institutes of Health) and shows the expression levels of ezrin and NHE3 normalized to vinculin in kidney fractions. For ezrin, only the upper bands, which represent ezrin, were quantified. Similar results were obtained at least seven times for BBM prepared from either male or female mice. (C–F) The expression of phosphorylated ezrin was analyzed with phospho-ERM antibody by immunofluorescence of kidney (C and D) and jejunum (E and F) sections from EBP50(+/+) (C and E) and (–/–) (D and F) 5-week-old littermates. The samples were imaged with a Zeiss LSM 510 confocal microscope by using the HeNe 543-nm laser and identical imaging parameters. Note the higher apical expression of phosphorylated ezrin (arrows) in wild-type kidney tubules and intestinal villi.