Cellular localization of mCherry-tagged SAMHD1 and SAMHD1 mutant proteins using live cell imaging.

<p>(A)–(F) Plasmid pEYFP-Nuc (Clontech) and pmCherry-SAMHD1 WT or indicated variants were co-transfected into HEK293T cells using PEI Max (Polysciences). For live cell imaging, HEK293T cells were transfected in 6-well coverslip glass-bottomed cell culture dishes (InVitro Scientific) when the cells were ∼80% confluent, and then visualized after 24 h using a Zeiss LSM510-Meta confocal imaging system equipped with four argon lasers (458-, 477-, 488-, and 514-nm lines), two HeNe lasers (542 and 633 nm), and one diode laser (405 nm). All images were acquired from a 100X objective, and image analysis and manipulation were performed using Zen 2009 software.