A Hsp40 Chaperone Protein Interacts with and Modulates the Cellular Distribution of the Primase Protein of Human Cytomegalovirus
Figure 5
Up- and down-regulated expression of DNAJB6 in cells.
Western blot analysis of the levels of DNAJB6a (lanes 1–4) and b (lanes 5–8) and Myc-tagged UL70 in the parental U251 cells (U251) (lanes 1 and 5), the DNAJB6a-expressing U251-6a (U251-6a) (lane 4) and DNAJB6b-expressing cells (U251-6b) (lane 8), or U251 cells that were transfected with anti-DNAJB6a siRNA (6a-siRNA) (lane 3), anti-DNAJB6b siRNA (6b-siRNA) (lane 7), or control siRNA (C-siRNA) (lanes 2 and 6). The expression of cellular actin was used as the internal control. Cells were transfected with pCMV-Myc-UL70 in the absence and presence of siRNAs. Forty-eight hours after transfection, cells were infected with HCMV at MOI of 1. Protein samples were prepared at 48–72 hours postinfection. The membranes were reacted with antibodies, stained using a Western chemiluminescent substrate kit (GE Healthcare), and quantitated with a STORM840 PhosphorImager (GE Healthcare) or a Gel Documentation Station (BioRad, Hercules, CA) [54], [55]. A dilution series of the samples was analyzed and the results were compared in order to accurately determine the protein levels. Quantitation was performed in the linear range of protein detection [54], [55].