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. 2019 Jun 5;10(1):186–194. doi: 10.1016/j.apsb.2019.05.005

Figure 3.

Figure 3

IGF-1 regulates OAT3 expression through PKA pathway. (A) Top panel: OAT3-expressing cells were treated with IGF-1 (100 nmol/L) with or without PKA inhibitor H89 (10 μmol/L) for 4 h. Cells were labeled with membrane impermeable biotin. Biotinylated cell surface proteins were separated with streptavidin beads, followed by immunoblotting (IB) with anti-myc antibody (OAT3 was tagged with epitope myc for immunodetection). Bottom panel: The identical blot of the top panel was re-probed with anti-E-cadherin antibody. E-cadherin is a marker for cell membrane proteins. (B) Densitometry analysis of blot results from Fig. 3A top panel as well as from other experiments. The values are mean ± SD (n = 3); *P < 0.05. (C). Top panel: OAT3-expressing cells were treated with IGF-1 (100 nmol/L) with or without H89 (10 μmol/L) for 4 h. The cells were collected and lysed, followed by immunoblotting with anti-myc antibody. Bottom panel: The identical blot of the top panel was re-probed with anti-GAPDH antibody. GAPDH is a marker for total cell proteins. (D). Densitometry analyses of blot results from Fig. 3C top panel as well as from other experiments. The values are mean ± SD (n = 3); *P < 0.05. Statistical analysis was performed using one-way ANOVA by using GraphPad Prism software (GraphPad Software Inc., San Diego, CA, USA).