MNDAL protein expression. (A) Western blot analysis of cells transfected with V5-His–tagged IFI205 (detected at ∼ 50 kDa) and MNDAL (detected at ∼ 60 kDa). (B-C) Immunofluorescent staining of NIH3T3 cells transfected with V5-His–tagged IFI205 (B) and XRPC24 PCT cells transfected with V5-His–tagged MNDAL (C). Staining with V5 antibody (green) showed overlap with DAPI (blue) stained nuclei. Cells were examined at 25°C with a Zeiss LSM 510 NLO confocal system (Carl Zeiss Inc) with an Axiovert 200M inverted microscope and operating with a 2-photon laser tuned to 750 nm, 25 mW argon laser tuned to 488 nm, and 1 mW HeNe laser tuned to 543 nm. Cells were imaged with a 63×/1.4 NA Zeiss Plan-Apochromat oil-immersion objective. Digital images (512 × 512 pixels, 8 bit) were collected with the use of the Zeiss AIM software with a scan zoom from 2 to 4 and a multitrack configuration in which the FITC, Cy3, and DAPI signals were collected sequentially with a BP 500- to 530-nm filter, with BP 565- to 615-nm filter, and BP 390- to 465-nm filter after excitation with 488-nm, 543-nm, and 750-nm laser lines, respectively.