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Blood, Vol. 104, Issue 9, 2736-2738, November 1, 2004
Susceptibility of human fetal mesencyhmal stem cells to Kaposi sarcoma-associated herpesvirus
Blood Parsons et al.
104: 2736
Supplemental materials for: Parsons et al, Vol 104, Issue 9, 2736-2738
Files in this Data Supplement:
- Figure 1. KSHV-infected MSC cultures display only low levels of lytic gene expression (JPG, 16 KB)
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Immunofluorescence assays using antibodies to KSHV lytic gene products were performed 72 hours after 12-O-tetradecanoyl phorbol-beta-acetate (TPA) and sodium butyrate induction of MSC that had been infected one day earlier with cell-free KSHV (see "Materials and methods"). Nuclear staining with DAPI (panels A, C, E, and G) is compared with images of rare cells expressing lytic proteins (see "Results and discussion"). Panels B, D, F and H are merged images combining antibody staining results for LANA (green) and lytic proteins (red) for MCP (B), RTA/ORF50 (D), MIR-1/K3 (F), and MIR-2/K5 (H). Only RTA (D) was detectable by IFA at an appreciable frequency (8% of induced cells); expression of the other lytic proteins tested, though present, was rare (< 0.08% of induced cells). Neither latent nor lytic antibody staining was present in control cells cultured with ultraviolet (UV)?inactivated KSHV or with infected cells employing secondary antibodies alone (data not shown). Microscopy and photography were performed using a Nikon Eclipse TE2000-E fluorescence microscope with attached Nikon digital camera. Original microscopy employed a 40X objective. Acquisition and processing of photographs were performed using Improvision software (Lexington, MA).
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