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. 2008 Jun 6;283(23):15799–15806. doi: 10.1074/jbc.M801553200

FIGURE 7.

FIGURE 7.

Surface TrkB expression regulated by D1 receptor agonist. Striatal cultures were treated with control vehicle (PBS), SKF38393 (SKF), and BDNF on DIV6. Some of the cultures were preincubated with SCH23390 (A, SCH+SKF), K-252a (A), BAPTA (B, BAPTA+SKF), or BAPTA-AM (B, BAPTA-AM+SKF) in 37 °C prior to SKF38393. The cultures were incubated with biotin-PBS for 30 min on ice and then lysed with radioimmune precipitation assay buffer. Half of biotinylated proteins were precipitated by avidin-conjugated beads and then subjected to SDS-PAGE and Western blotting for TrkB. The other samples were used for immunoprecipitation (IP) with TrkB. Duplicate samples are displayed. Typical immunoblots (IB) are shown. Protein levels were determined from independent wells (n = 4). Quantifications of surface biotinylated TrkB levels by NIH Image are shown. *, p < 0.05. The bars indicate S.D. C–H, striatal cultures were treated with control (PBS) and SKF38393 on DIV6. Some of the cultures were preincubated with SCH23390 (SCH+SKF), BAPTA (BAPTA+SKF), or BAPTA-AM (AM+SKF) in 37 °C prior to SKF38393. The cultures were fixed and immunostained with rabbit control IgG (C) or TrkB (N terminus) antibody (D–H) without detergent. Representative pictures are provided.