Figure 2. Expression of mutant NHS-A proteins in mammalian cells. A: Lysates of HEK 293A cells transiently transfected with GFP-NHS-A400delC and pEGFP-C1 constructs and untransfected cells were analyzed by western blotting with anti-GFP antibody. B: Lysates of HEK 293A cells transiently transfected with FLAG-NHS-A, FLAG-NHS-AC2701T and FLAG-NHS-A4071del299bp in pCMV-Tag 2A and untransfected cells were analyzed by western blotting with anti-FLAG tag antibody. A protein band of greater than 150 kDa seen in all the lanes is due to non-specific binding of the anti-FLAG tag antibody (indicated with an asterisk). A very faint protein band of approximately 130 kDa in the NHS-A and 4071Δ299bp lanes is most likely due to protein degradation. The molecular masses of proteins standards are indicated. UT=untransfected cells.