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Fig. 5. Endogenous SCAMPs bind and largely colocalize with NHE7. (A) Control PC12 cells (PC12) or PC12 cells stably expressing 1D4-tagged NHE7 (NHE7/PC12) were immunoprecipitated (IP) with the 1D4 antibody conjugated to sepharose beads. Co-immunoprecipitated SCAMP1, 2, or 5 was detected by western blot probed with anti-SCAMP antibodies. (B) Fixed NHE7/PC12 cells on glass coverslips were double stained with anti-SCAMP1, SCAMP2 or SCAMP5 rabbit antibody with anti-1D4 mouse antibody. SCAMP1 (a), SCAMP2 (d) and SCAMP5 (g) were visualized with Alexa 488-conjugated goat anti-rabbit IgG (green) and the corresponding NHE7 by Alexa 568-conjugated anti-mouse IgG (red fluorescence) (b, e and h respectively). Yellow signals in merged images (c, f and i respectively) correspond to colocalized proteins. Bar, 10 µm.





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