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. 2008 May 9;283(19):13216–13224. doi: 10.1074/jbc.M706749200

FIGURE 6.

FIGURE 6.

5HT and 5HTP production in PC12 cells. PC12 cells were transfected with wild-type HA-TPH2, HA-TPH2 S19A, or HA-TPH2 S19D, and 48–72 h post-transfection, 5HT was measured, or cells were treated with 100 μm NSD-1015 to block the activity of AADC and either 2.5 μm forskolin or an equal volume of DMSO for 16 h to assess accumulation of 5HTP. A, 5HT content in untreated cells was measured by HPLC and normalized to the endogenously produced dopamine content produced by the cells to control for differences in the number of cells used to produce the lysates. ‡, p < 0.05 compared with HA-TPH2; †, p < 0.05 compared with S19A. B, similar to the results in A, 5HTP content was measured in cells transfected with each of the three constructs and treated with 100 μm NSD-1015 for 16 h to block AADC as described above (*, p < 0.001 compared with HA-TPH2). 5HTP accumulation was also assessed in cells treated with forskolin and NSD-1015 for 16 h (**, p < 0.001 compared with untreated controls; %, p < 0.05 compared with HA-TPH2 + forskolin; #, p < 0.001 compared with HA-TPH2 + forskolin). 5HTP content in the cell lysates was also normalized to the endogenously produced dopamine content produced by the cells before the initiation of treatment to control for differences in cell number used to produce the lysates. Data are means ± S.E. (n = 4).