Supporting Online Material


Abstract
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Cytomegalovirus Recruitment of Cellular Kinases to Dissolve the Nuclear Lamina
Walter Muranyi, Jürgen Haas, Markus Wagner, Georg Krohne, and Ulrich H. Koszinowski

Supporting Online Material

Supplemental Figure 1. Ca-dependent PKCs are specifically recruited to the NE in MCMV infected cells. Ca-dependent PKCs (A-C), but not MAPK (D-F), protein kinase A RI subunit (G-I) or casein kinase II (J-L) are recruited to the INM in NIH 3T3 fibroblasts infected with MCMV. Cells were stained with antibodies against protein kinases and either pp89IE1 or M50/p35 as a control for infection.


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Supplemental Figure 2. MCMV infection induces a subcellular distribution of PKC to the NE. NIH 3T3 infected with MCMV at a MOI of 0.5 were analyzed 6 hours (A, D), 16 hours (start of virus morphogenesis) (B, E), and 20 hours (peak of virus morphogenesis, first infectious progeny) after infection (C, F). Cells were stained with antibodies to PKC and the nuclear immediate early protein 1 (pp89IE1) as a marker for infected cells. PKC shows a diffuse staining in the cytosol as well as a dotted staining at the plasma membrane at 6h p.i. After 16 and 20h p.i., PKC is localized at the NE and at 20h p.i. additional dotted structures appear at the NE.


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Supplemental Figure 3. M50/p35 contains a LEM-like domain. (A) M50/p35 contains a LEM-like domain between amino acid residues 190 to 243. Hydrophilicity blot and schematic diagram of M50/p38 indicating the location of the LEM-like domain (open box) and the transmembrane region (filled box). (B) M50/p35 is homologous to INM proteins containing a LEM domain. Sequence comparison between M50/p35, Emerin, MAN1, the LEM-like (LAP2Greek Letter Beta-N) and the LEM domain (LAP2Greek Letter Beta-C) of LAP-2Greek Letter Beta?. The numbers indicate the amino acids residues of the protein sequence shown. Identical amino acids are marked in dark grey, similar amino acids in light grey.


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