Figure 1.

Gα₁₂ interacts with JLP. (A) COS-7 cells were cotransfected with vectors encoding S-JLP along with wild-type form (Gα₁₂WT) or constitutively active form (Gα₁₂QL) of Gα₁₂ expression constructs. Lysates were collected after 48 hours, and the pull-down assay was carried out using S-agarose beads. Immunoblot analysis was carried out to assess the presence of Gα₁₂ in the S-JLP precipitates, using an antibody specific to Gα₁₂. Expression levels of JLP and Gα₁₂ were monitored by immunoblot analysis with Gα₁₂ or S-protein antibody. Results form a typical experiment presented here (n = 3). (B) COS-7 cells were cotransfected with vectors encoding S-JLP along with wild-type form (Gα₁₂WT). Transfectants were serum-starved for 24 hours after which the cells were stimulated with 20 µM of LPA for 10 minutes. After LPA stimulation, cells were lysed, and the lysates were subjected to S-agarose pull-down assay. Immunoblot analysis was carried out to assess the presence of Gα₁₂ in the S-JLP precipitates, using an antibody specific to Gα₁₂. Expression levels of JLP and Gα₁₂ were monitored by immunoblot analysis with Gα₁₂ or S-protein antibody. Results form a typical experiment from a set of three repeats.