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. Author manuscript; available in PMC: 2015 Apr 6.
Published in final edited form as: Regul Pept. 2007 Jun 21;144(0):72–81. doi: 10.1016/j.regpep.2007.06.004

Table 1.

Long-term inhibition of Y2 receptor and G-protein activity in CHO cells by pertussis toxin

Group Y2 binding basal GTP- γ-S binding PYY- stimulated GTP-γ-S binding % decrease of Y2 binding % decrease of basal GTP-γ-S % decrease of PYY- stimulated GTP-γ-S
Control 272 ± 1.08 84.2 ± 0.11 54.3 ± 0.23
PTX no recovery 51.8 ± 0.84 35.7 ± 1.2 6.53 ± 0.98 81 ± 1.3 57.6 ± 1.9 88 ± 13.2
PTX 2 d recovery 50.5 ±0.31 27.3 ± 1.19 4.33 ± 0.77 81.4 ± 0.5 67.6 ± 3 92 ± 16.4
PTX 7 d recovery 41.4 ± 0.03 30.8 ± 0.93 7.86 ± 0.68 84.8 ± 0.06 63.4 ± 1.9 85.5 ± 7.4
PTX pass 1 – 11 d recovery 37.4 ± 3.65 37 ±0.71 3.55 ± 0.62 86.3 ± 8.4 56.1 ± 1.1 93.5 ± 16.3
PTX pass 2 – 14 d recovery 30.8 ± 0.27 37.6 ±4.6 6.93 ±4.08 88.7 ± 0.78 55.3 ± 6.7 87.2 ± 12.8

The cells were incubated with PTX (4 ng/ml growth medium) for 24 h, washed, the medium replaced and cells then collected immediately (no recovery) and 2 or 7 days later. Passage 1 was done on recovery day 7, and passage 2 on recovery day 11. Cells from that passage were collected on recovery day 14. Particulates from all samples were assayed for the binding of 50 pM of [125I]PYY(3-36) and 200 pM of [35S]GTP-γ-S (basal and stimulated by 100 nM PYY), as specified in Methods. All binding results are in fmol/mg particulate protein. The Y2 agonist –stimulated GTP-γ-S binding is the increase over the basal. All decrease percentages are relative to control. Cells from four wells were assayed for all PTX-treated groups, and cells from eight wells for the control group. Data are shown ± 1 S.E.M. .