. 2021 Feb 20;46(1):29–42. doi: 10.1584/jpestics.D20-079

Table 1. In vitro RGA systems for the screening of EcR-USP ligands.

Host cells	Organism and cell lines	Transfected receptor genes	EcRE	Reporter gene	Types of test compounds	Ref.
Insect cells*	Drosophila melanogaster Kc	—	Dm hsp27 promoter	Luc	20E, α-ecdysone	55)
	D. melanogaster Kc	—	Dm hsp27 promoter	Luc	3,5-di-tert-butyl-4-hydroxy-N-isobutyl-benzamide (DTBHIB)	56)
	D. melanogaster Kc	—	Dm hsp27 promoter	Luc	Diacylhydrazine (DAH) compounds	57)
	D. melanogaster Schneider 2 (S2)	—	Dm hsp27	lacZ	Cucurbitacins	134)
	D. melanogaster Schneider 2 (S2)	—	EcRE	lacZ	Cucurbitacins	134)
	D. melanogaster Schneider SL2	—	Dm hsp27	Luc	Insect and plant ecdysteroids	139)
	D. melanogaster Schneider SL2	—	EcRE	Luc	Insect and plant ecdysteroids	139)
	Spodoptera frugiperda Sf9	—	Dm hsp27	Luc	20E, ponasterone A (pon A), chromafenozide	67)
	Spodoptera frugiperda Sf9	—	EcRE	Luc	20E, ponasterone A (pon A), chromafenozide	67)
	D. melanogaster Kc	—	Dm hsp27	Luc	20E, pon A, chromafenozide	67)
	D. melanogaster Kc	—	EcRE	Luc	20E, pon A, chromafenozide	67)
	Bombyx mori Bm5	—	Dm hsp27	CAT	Plant compounds	58)
	Bombyx mori Bm5	—	EcRE	GFP	DAH library	58)
	S. frugiperda Sf9	CfEcR	Dm hsp27	CAT	RG-102240	68)
	S. frugiperda Sf9	CfEcR	EcRE	CAT	RG-102240	68)
	Lymantria dispar (Ld652Y)	CfEcR	GAL4 RE (two-hybrid)	CAT	RG-102240	68)
	Lymantria dispar (Ld652Y)	MmRXR	GAL4 RE (two-hybrid)	GFP	RG-102240	68)
	B. mori Bm5	—	Dm hsp27	GFP	DAHs	70)
	B. mori Bm5	—	EcRE	GFP	DAHs	70)
	D. melanogaster S2	—	Dm hsp27	Luc	Chromafenozide	69)
	D. melanogaster S2	—	EcRE	Luc	Methoxyfenozide	69)
	B. mori Bm5	—	Dm hsp27	Luc	Chromafenozide	69)
	B. mori Bm5	—	EcRE	Luc	Methoxyfenozide	69)
	Anthonomus grandis BRL-AG-3A	—	Dm hsp27	Luc	DAHs	76)
	BRL-AG-3C	—	EcRE	GFP	DAHs	76)
	Leptinotarsa decemlineata BCRL-Lepd-SL1	—	Dm hsp27	Luc	DAHs	76)
	Leptinotarsa decemlineata BCRL-Lepd-SL1	—	EcRE	GFP	DAHs	76)
	D. melanogaster S2	—	Dm hsp27	CAT	DAHs, acylaminoketone (AAK) analogs, tetrahydroquinoline (THQ) compounds, piperidiamine analogs	80)
	D. melanogaster S2	—	EcRE	Luc		80)
	B. mori Bm5	—	Dm hsp27	GFP	DAHs, AAK analogs, THQs	71)
	B. mori Bm5	—	EcRE	GFP	DAHs, AAK analogs, THQs	71)
	S. littoralis Sl2, Slj2b	—	Dm hsp27	Luc	DAHs, AAK analogs, THQs	71)
	S. littoralis Sl2, Slj2b	—	EcRE	Luc	DAHs, AAK analogs, THQs	71)
	S. frugiperda Sf9	—	Dm hsp27	Luc	Non-steroidal compounds	78)
	S. frugiperda Sf9	—	EcRE	Luc	Non-steroidal compounds	78)
	L. decemlineata	—	Dm hsp27	Luc	DAHs	77)
	BCRL-Lepd-SL1	—	EcRE	Luc	DAHs	77)
	D. melanogaster S2	—	Dm hsp27	Luc	Ecdysteroids	135)
	D. melanogaster S2	—	EcRE	Luc	Ecdysteroids	135)
	B. mori Bm5	—	Dm hsp27	Luc	Ecdysteroids	135)
	B. mori Bm5	—	EcRE	Luc	Ecdysteroids	135)
	S. frugiperda Sf9	—	Dm hsp27	Luc	N-tert-butylphenyl thenoylhydrazide compounds	140)
	S. frugiperda Sf9	—	EcRE	Luc	N-tert-butylphenyl thenoylhydrazide compounds	140)
	D. melanogaster S2	—	Dm hsp27	Luc	Compounds registered in Maybridge database	79)
	D. melanogaster S2	—	EcRE	Luc	Compounds registered in Maybridge database	79)
	B. mori Bm5	—	Dm hsp27	Luc	Compounds registered in Maybridge database	79)
	B. mori Bm5	—	EcRE	Luc	Compounds registered in Maybridge database	79)
Mammalian cells	Cricetulus griseus	VgEcR	E/GRE	lacZ	Phytochemicals	90)
	CHO	HsRXR	E/GRE	lacZ	Phytochemicals	90)
	Mus musculus	AaEcR	GAL4 RE (two-hybrid)	Luc	THQs	3)
	NIH 3T3	MmRXR	GAL4 RE (two-hybrid)	Luc	THQs	3)
Mammalian cells	Homo sapiens	BmEcR	Dm hsp27 EcRE	β-gal	AAKs	72)
	HEK293	BmEcR		β-gal	AAKs	72)
	H. sapiens	BmEcR	Dm hsp27 EcRE	β-gal	AAKs	60)
	HEK293	BmEcR		β-gal	AAKs	60)
	C. griseus	CfEcR	GAL4 RE (two-hybrid)	Luc	AAKs	60)
	CHO	HsRXR-LmRXR chimera	GAL4 RE (two-hybrid)	Luc	AAKs	60)
	M. musculus	CfEcR	GAL4 RE (two-hybrid)	Luc	DAHs, THQs	61)
	NIH 3T3	LmRXR
	NIH 3T3	DmTai
	M. musculus	AaEcR	GAL4 RE (two-hybrid)	Luc	THQs	4)
	NIH 3T3	CfEcR
		DmEcR
		MsEcR
		BmEcR
		LmEcR
		TmEcR
		AmaEcR
		HsRXR
	M. musculus	AaEcR	GAL4 RE (two-hybrid)	Luc	Natural and semi-synthetic ecdysteroids (hydroxylation)	62)
	NIH 3T3	CfEcR
		DmEcR
		MsEcR
		BmEcR
		TmEcR
		AmaEcR
		BaEcR
		NcEcR
		HsRXR
	M. musculus	VgEcR	E/GRE	Luc	Natural and semi-synthetic ecdysteroids (hydroxylation)	62)
	NIH 3T3	HsRXR	E/GRE	Luc	Natural and semi-synthetic ecdysteroids (hydroxylation)	62)
	M. musculus	AaEcR	GAL4 RE (two-hybrid)	Luc	Natural and semi-synthetic ecdysteroids (alkylation)	63)
	NIH 3T3	CfEcR
	NIH 3T3	DmEcR HsRXR-LmRXR chimera
	M. musculus	VgEcR	E/GRE	Luc	Natural and semi-synthetic ecdysteroids (alkylation)	63)
	NIH 3T3	HsRXR	E/GRE	Luc	Natural and semi-synthetic ecdysteroids (alkylation)	63)
	H. sapiens	BmaEcR	GAL4 RE (two-hybrid)	Luc	Steroidal ecdysones, DAHs	64)
	HEK293	HsRXR	GAL4 RE (two-hybrid)	Luc	Steroidal ecdysones, DAHs	64)
Yeast	Saccharomyces cerevisiae	CfEcR-CfUSP	Dm hsp27 EcRE	β-gal	Nonsteroidal ecdysone agonists	85)
		HsRXR
		MmGRIP1
	S. cerevisiae	AaEcR-AaUSP	Dm hsp27 EcRE	β-gal	Ecdysteroids, nonsteroidal ecdysone agonists	84)
		DmUSP
		CfUSP
		HsRXR
		MmGRIP1
	S. cerevisiae	DmEcR-DmUSP	IR0	β-gal	Ecdysteroids, DAHs, THQ	87)
		CsEcR-CsUSP	DR2
		LdEcR-LdUSP	DR4
		DmTai	DR4

Abbreviations Aa: Aedes aegypti; Ama: Amblyomma americanum; Ba: Bemisia argentifolii; Bm: Bombyx mori; Bma: Brugia malayi; Cf: Choristoneura fumiferana; Cs: Chilo suppressalis; Dm: Drosophila melanogaster; E/GRE: ecdysone/glucocorticoid responsive element (hybrid RE); Hs: Homo sapiens; Lm: Locusta migratoria; Ms: Manduca sexta; Nc: Nephotettix cincticeps; Tc: Tribolium castaneum; Tm: Tenebrio molitor; Mm: Mus musculus; CHO; Chinese hamster ovary; GAL4 RE: DNA element for binding of the GAL4 DNA-binding domain (DBD); VgEcR: hybrid of D. melanogaster EcR carrying modified DBD and VP16 AD that recognizes an E/GRE *Reporter gene assays in insect cells expressing endogenous EcR and USP. Only the reporter plasmid was introduced.