Fig. 2
From: A loop region of BAFF controls B cell survival and regulates recognition by different inhibitors
Flap mutations affecting 60-mer formation: one of them additionally affects activity of BAFF 3-mer. Naturally cleaved, untagged human or mouse BAFF, with or without the indicated mutations in the flap, were recovered in supernatant of 293 T cells transiently transfected with plasmids encoding the full length wild type (WT) or mutant BAFF. Concentrated supernatants were fractionated by size-exclusion chromatography and fractions analyzed for the presence of BAFF by western blot using anti-human or anti-mouse BAFF antibodies, and for the activity of BAFF at the indicated dilutions of fractions using a hBAFFR:Fas reporter cell line that undergoes Fas-mediated killing upon stimulation of the chimeric receptor by BAFF. a Analysis of human BAFF WT and mutants. White arrows points at fractions with peak activity of BAFF 3-mer and 60-mer, as indicated. b Analysis of mouse BAFF WT and mutants. The experiment was performed twice for human BAFF and thrice for mouse BAFF
