Fig. 2
The effect of PKC on OAT1 expression. (A, top panel): cell surface expression of OAT1. OAT1-expressing COS-7 cells were treated for 1 h, with 10 μM PMA in the absence or presence of 2 μM staurosporine (St). Cells were then labeled with membrane-impermeable biotinylation reagent. Biotinylated cell surface proteins were then separated by streptavidin resin, followed by immunoblotting (IB) with anti-myc antibody (epitope myc was tagged to OAT1). (A, Bottom panel): The same immunoblot from the top panel was stripped and reprobed with anti-E-cadherin antibody to determine the expression of the cell surface protein marker E-cadherin. (B): Densitometry plot of results from (A) as well as from other repeat experiments. The values are mean ± S.D. (n = 3). *p < 0.05. (C, top panel): cell total expression of OAT1. (C, bottom panel): The same immunoblot from the top panel was stripped and reprobed with anti-β-actin antibody to determine the expression of β-actin, a protein marker for total cellular proteins. (D): Densitometry plot of results from (C) as well as from other repeat experiments. The values are mean ± S.D. (n = 3)