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. Author manuscript; available in PMC: 2009 Aug 31.
Published in final edited form as: J Biol Chem. 2006 Feb 16;281(16):11097–11103. doi: 10.1074/jbc.M511460200

FIGURE 2. Effect of transient expression of Rab4 on the recycling of internalized β-AR in HL-1 myocytes.

FIGURE 2

A, immunoblot analysis of Rab4 expression. HL-1 myocytes were transiently transfected with FLAG-tagged Rab4 or the pcDNA3 vector (Control). Rab4 expression was determined by Western blotting using anti-FLAG (upper panel) and Rab4 antibodies (lower panel). Rab4 antibodies detected both exogenous and endogenous Rab4. B, effect of Rab4 on the recycling of ISO-mediated internalized β-AR. HL-1 cells were transiently transfected for 48 h with Rab4 (squares) or the pcDNA3 vector (triangles), stimulated with ISO (10 μM) for 30 min, and allowed to recover for 15, 30, 60, 120, and 240 min at 37 °C. Cell surface expression of β-AR was measured by ligand binding as described in the legend of Fig. 1. β-AR expression at the cell surface are 2382 ± 212 cpm in the absence of ISO and 952 ± 317 cpm after 30 min exposure to ISO. The data shown are the percentage of β-AR recycling in the cell transfected with pcDNA3 and recovered for 240 min after ISO stimulation and presented as the means ± S.E. of six independent experiments. *, p < 0.05 versus control at the same time points.