Analysis of the effects of pyrin domain deletions on colocalization with microtubules.

(A) Cos-7 cells were transfected with plasmids encoding GFP-pyrin deleted for one or more domains. Cells were treated with 10 μM paclitaxel before fixing. Microtubules were detected with mouse anti–β-tubulin antibody and rhodamine-labeled secondary antibody. FL indicates full-length pyrin; N, amino acids 1 to 374 of pyrin (N-terminal domain); NB, amino acids 1 to 407 of pyrin (N-terminal plus B-box zinc finger); NBC, amino acids 1 to 576 of pyrin (N-terminal, B-box zinc finger and coiled-coil); NBR, amino acids 1 to 407, 577 to 781 of pyrin (N-terminal, B-box zinc finger, RFP domain); NCR, amino acids 1 to 374, 408 to 781 of pyrin (N-terminal, coiled-coil, RFP); BCR, amino acids 374 to 781 of pyrin (B-box zinc finger, coiled-coil, RFP); R, amino acids 577 to 781 of pyrin (RFP domain). (B) Schematic of deletion/truncation series. (C) NBC construct with altered microtubule appearance shows colocalization of microtubules (upper panel) with GFP signal (lower panel). Magnification × 1000.