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. 2007 May;18(5):1850–1860. doi: 10.1091/mbc.E06-08-0765

Figure 7.

Figure 7.

Requirement of both R-Ras and RalA for depolarization-induced exocytosis. (A) PC12 cells were transfected with pVenus-NPY together with the indicated constructs. Sixty hours after transfection, the cells were stimulated with high-potassium saline for 20 min. The efficiency of NPY-Venus secretion was determined as the ratio of NPY-Venus present in the medium versus that remaining in the cell lysates. The values were normalized to a vector control. Error bars indicate the SD from at least three experiments. The symbols indicate the results of t test analysis; *p < 0.002 compared with the control. (B and C) PC12 cells were transfected with pVenus-NPY and siRNA for luciferase, R-Ras, RalA, RalB, RalGDS, Rgl, or Rgl2/Rlf. NPY secretion upon depolarization was examined as described in A. Error bars indicate the SD from at least three experiments. The symbols indicate the results of t test analysis; *p < 0.001 compared with the control. (D) GH3 cells and GH3/ R-Ras cells were transfected with expression vectors for NPY-Venus or EYFP-GH1. Depolarization-induced secretion was monitored as described in A. The symbols indicate the results of t test analysis; *p < 0.001 compared with the control. (E) GH3/R-Ras cells were transfected with expression vectors for NPY-Venus and an shRNA vector for R-Ras, or RalA. Depolarization-induced secretion of NPY was examined as described in A. The symbols indicate the results of t test analysis; **p < 0.001 compared with the control (GH3/R-Ras cells).