Table 2.

Site-directed mutagenesis primers for HA-RXFP3 D/E to A mutants.

Mutation	Direction	Primer sequence (5′–3′)
E47A/D51A	Fwd	CCG GCC TTG TGG TGG GCG CTG GGG C
	Rev	CAG CGC CCA CCA CAA GCC CGG AAG CTG
E55A/D58A	Fwd	CTG GCC TTG CCG GCC GGC GCG CCG CC
	Rev	GCC GGC CGG CAA GGC CAG CCC CAG CTC CCA CC
E72A	Fwd	GCA GCG AGC GCG GAC ACA GAG GCC C
	Rev	GCT CGC TGC CCC GCC GCT G
D75A	Fwd	GCG GCC ACA GAG GCC CGG GTG C
	Rev	C TGT GGC CGC GCT CTC TGC CCC
E77A	Fwd	CACA GCG GCC CGG GTG CGG ATT CTC ATC
	Rev	GGC CGC TGT GTC CGC GCT CTC TGC CC
E141A/D145A	Fwd	GTG GCC AAC GCT CTT GCC TTC AAA TGG CCC TTC GGC AAG G
	Rev	GAA GGC AAG AGC GTT GGC CAC CGC CCA GAA GGG CAG
E141A	Fwd	GTG GCC AAC GCT CTT GAC TTC AAA TGG CCC TTC
	Rev	GTT GGC CAC CGC CCA GAA GGG CAG
D145A	Fwd	CTT GCC TTC AAA TGG CCC TTC GGC AAG GCC
	Rev	GAA GGC AAG AGC GTT CTC CAC CGC CCA G
E244A	Fwd	GGC GCC GAG CTG TGC CTG GTG CGT TTC
	Rev	CTC GGC GCC CAT CAC CTT GAC CGT GG
E245A	Fwd	GAG GCC CTG TGC CTG GTG CGT TTC C
	Rev	CAG GGC CTC GCC CAT CAC CTT GAC CG
E253A	Fwd	CCG GCC AAG TTG CTG GGC CGC G
	Rev	CTT GGC CGG GAA ACG CAC CAG GC
D259A	Fwd	CGC GCC AGG CAG TTC TGG CTG GGC CTC
	Rev	CCT GGC GCG GCC CAG CAA CTT GTC CG
E362A	Fwd	CAG GCG ATT TTCC TGT GCC AGG TAT ACG CGT TC
	Rev	GAA ATA CGC CTG GCT GAA GGG CAC CGC G