Fig 1: Activated CRLs containing CUL1 and especially CUL2 and CUL3 regulate IL-2 production.(A) MA5.8ζ IL-2 secretion at 12 h normalized to the number of live cells measured by annexin V and propidium iodide staining and (B) transcription at 12 h with 4 µg/mL α-CD3 and the indicated concentrations of OPT (n = 3). (C–G) IL-2 secretion by MA5.8ζ cells, cells expressing control shRNA, and cells expressing an shRNA for (C) CUL1, (D) CUL2, (E) CUL3, (F) CUL4a/b and (G) CUL5 after 48 h of stimulation with indicated concentrations of α-CD3. Data are averaged from two independent experiments, where each experiment examined three independently created lines for each shRNA. (H) IL-2 secretion at 48 h by MA5.8ζ cells and cells expressing either control shRNA, CUL2 shRNA or CUL3 shRNA with 1 µg/mL α-CD3 and 200 nM MLN4924. Data are averaged as in (C–G). Values represent mean ± s.e.m.; *P<0.05, **P<0.01, ***P<0.001.