Figure 3
From: Characterization and identification of PARM-1 as a new potential oncogene
PARM-1 protein profile and secretion by NIH/3T3 cells. Immunoblotting of lysates (30 μg) from NIH/3T3 cells transiently transfected with expression vector of hParm-1 (full-length or mutants) or of mParm-1 using (a) an anti-hPARM-1 (Sigma; 1:1000) antibody or (b) an anti-GFP (Santa Cruz Biotechnology; 1:1000) antibody. Culture supernatants from these cells were collected, centrifuged, concentrated and subjected to SDS-PAGE (12 %) and hPARM-1 protein was detected by western blot (a) using anti-hPARM-1 and (b) anti-GFP antibodies recognizing respectively the N-terminus and C-terminus of hPARM-1 fusion protein. The region below 80-kDa from the supernatant in panels a and b contains no detectable bands. The anti-β-actin (Sigma; 1:1000) was used to detect possible media contaminations by proteins from lysed cell. (c) mPARM-1 expression was also tested in cell lysates.