Western blot analysis of the production of IL-12Rβ1 chains.

(A) Cell lysates from various cells were subjected to Western blot analysis using an antibody against the carboxy terminus of human IL-12Rβ1 protein (polyclonal), and the production of an IL-12Rβ1 chain protein of approximately 100 kd was examined. Lane 1, HEKC293 transfected with wild-type IL-12Rβ1wt gene; lane 2: HEKC293 transfected with mutated IL-12Rβ1R213W gene; lane 3, HEKC293 mock transfected with an expression vector only; lane 4, TS-1HTLV-1 cells; lane 5, cells from a healthy subject. The lower band represents a nonspecific band. (B) Cell lysates were similarly treated, but a different anti–IL-12Rβ1 antibody (polyclonal) was used to confirm the authenticity of the 100-kd band visualized. Lane 1, HEKC293 cells transfected with an expression vector carrying the wild-typeIL-12Rβ1wt gene; lane 2, HEKC293 cells transfected with expression vector carrying a mutated IL-12Rβ1 gene; lane 3, HEKC293 cells transfected with expression vector only (mock); lane 4, TS-1HTLV-1 cells, an HTLV-I–immortalized T-cell line derived from the index patient; lane 5, PHA-PBMCs from the patient; lane 6, PHA-PBMCs from a healthy subject. Note that the same 100-kd protein profile was seen in panels A and B.