An Epstein-Barr virus that expresses only the first 231 LMP1 amino acids efficiently initiates primary B-lymphocyte growth transformation - PubMed (original) (raw)

FIG. 2

(Top) Immunoblot analysis of LCLs for LMP1. LMP1 was detected with S12 (36) monoclonal antibody from LCLs infected with WT EBV but not from LCLs infected with MS231 virus, since the S12 recognition sequence is not expressed in MS231. Lanes WT3, WT4, and WT8 contained protein from WT LCLs, and lanes 0, 1, 2, 3, 4, 6, and 7 contained protein from LCLs MS231-0, MS231-1, MS231-2, MS231-3, MS231-4, MS231-6, and MS231-7, respectively. BJAB is an EBV-negative B-lymphoma cell line. B95-8 is a WT EBV-infected marmoset cell line. Proteins from 2.5 × 105 cells were separated by SDS–8% PAGE. Size markers (kilodaltons) are indicated at the left. A truncated form of LMP1 is also expressed in B95-8 due to lytic replication occurring in this cell line (21). S12 monoclonal antibody was detected with sheep anti-mouse immunoglobulin conjugated to horseradish peroxidase (Amersham), followed by a chemiluminescence assay. (Middle) Immunofluorescent staining with rabbit antiserum recognizing the LMP1 amino terminus (22). (A) LCL infected with MS231 virus. (B) LCL infected with WT EBV. Magnification, ×1,000. (Bottom) Immunoblot analysis of LCLs for EBNA proteins. The EBNA 1, 2, 3, and LP proteins were detected with human serum and are indicated on the right. EBNA LP has multiple sizes in each LCL due to variable numbers of a repeat element within the gene. Lanes WT3, WT4, and WT8 contained protein from WT LCLs. Lanes 0, 1, 2, 3, 4, 6, and 7 contained protein from LCLs MS231-0, MS231-1, MS231-2, MS231-3, MS231-4, MS231-6, and MS231-7, respectively. Proteins from 5 × 105 cells were loaded in each lane. EBNA proteins were detected with human serum and protein A conjugated to horseradish peroxidase (Amersham), followed by chemiluminescence assay. Size markers (kilodaltons) are indicated on the left.