Because sRAGE scavenges RAGE ligands and reduces RAGE signaling
to NF-jB transcription program [35], we next tested
whether sRAGE produced from OP-transfected cells blocks NF-jB
activity as efficient as that produced in WT-transfected cells. We
found that sRAGE produced in OP-transfected cells blocked RAGE
ligands, HMGB1-, S100B-, and AGE-induced NF-jB activation, and
that the blocking efficiency is similar to that of sRAGE produced
in WT-transfected cells (Fig. 7). Given that our recent studies demonstrating
the key role of N-glycosylation on sRAGE bioactivity and
in vivo efficacy [17], and that the specific N-glycoform modification
may not be sufficiently resolved in SDS–PAGE (Fig. 6), the current
bioactivity assessment reaffirms that sRAGE produced from
OP-transfected cells is likely to be correctly modified, and therefore
is fully active. These assessments confirmed that sRAGE(OP) construct
boosts sRAGE production and can be used for future scaleup
production.