Identification of Caspase Cleavage Sites in KSHV Latency-Associated Nuclear Antigen and Their Effects on Caspase-Related Host Defense Responses
Fig 7
Mutation of both LANA caspase cleavage sites leads to increased IL-1β production.
THP-1 cells were matured overnight by treatment with TPA and then transiently transfected with FLAG-tagged forms of LANA-DM, WT-LANA, LANA-Nmut, or LANA-Cmut. The next day cells were treated with LPS and the level of IL-1β in the supernatants measured 20 hrs later. Cell extracts were also made for protein analysis and immunoblots. (A) IL-1β levels as determined by ELISA following transfection with the LANA plasmid constructs and treatment with LPS. The average level of IL-1β detected following transfection with vector alone (no LPS) was 2131 pg/ml (S3 Fig). (B) Immunoblot for cleaved caspase-1 and actin showing the relative levels of active casapse-1 compared to vector alone for the LPS-treated cells using the LiCor system. Data shown in (A) are the average +/- the standard deviation from 4 independent experiments. ** P< 0.01, *** P<0.005 for two-tailed Student’s t-test. Note that the average values for the single mutants were not statistically significantly different (p>0.05) when compared to wild type.