Figure 5.

The involvement of ERM and PIP₂ in the PPARα-induced NHE1 signaling pathway. (A) The interaction between NHE1 and ERM. NRK-52E cells were transfected with pcPPAR or blank vectors. NHE1 and ERM in transfected cells were immunoprecipitated with anti-NHE1 and anti-ERM antibodies, respectively. ERM and NHE1 in the immunocomplexes were detected by using Western blotting. IgG was detected as a loading control. Relative increases in the protein bands are also presented in bar chart form. Results are means ± SD (n = 3). *P < 0.05 by Student t test. (B) Interaction between ERM and PIP₂. ERM and PIP₂ in the transfected cells were immunoprecipitated with anti-ERM and anti-PIP₂ antibodies, respectively. PIP₂ and ERM in the immunocomplexes were detected by using Western blotting. Relative increases in the protein bands are also presented in bar chart form. Results are means ± SD (n = 3). *P < 0.05 by Student t test. (C) Influence of ERM siRNA transfection on apoptotic signals in the pcPPAR-transfected cells. The pcPPAR-transfected cells were further transfected with ezrin (Ez), radixin (Ra), moesin (Mo) or mock control (Mk) siRNA and then treated with or without 3 mmol/L gentamicin for 24 h. Total proteins of treated cells were analyzed by Western blotting with the specific antibodies as indicated. GAPDH was used as a loading control. Relative increases in the protein bands are also presented in a bar chart form. Results are means ± SD (n = 3). *P < 0.05 versus the mock control group. ^#P < 0.05 versus the mock control group with gentamicin treatment. Statistical differences between the two groups were determined using the Student t test.