Genotype	Measurement	Figure
behavior	wild type	+;+;+	Quantification of freezing and other behaviors	1C–J, S1B–E
	LC11 > shits	w−/+; R22H02-AD/UAS-shits; R20G06-DBD/UAS-shits	LC11 silencing	2A–G, S1F–H, S2, S3F
	empty / shits (split)	w−/+; BPp65-AD/UAS-shits; BPZp-DBD/UAS-shits	UAS control for synaptic silencing with split Gal4 drivers	2A–G, S1F–H, S2, S3B, D, G
	LC11 / +	w−/+; R22H02-AD/+; R20G06-DBD/+	Gal4 control for LC11 silencing and activation	2A–G, S1F–H, S2, S3F
	LC11 > ChR2-XXL	w−/+; R22H02-AD/UAS-ChR2-XXL; R20G06-DBD/+	LC11 optogenetic activation	2I–J
	empty / ChR2-XXL	w−/+; BPp65-AD/UAS-ChR2-XXL; BPZp-DBD/+	UAS control for optogenetic activation	2I–J
	LC10 > shits	w−/+; +/UAS-shits; R22D06-Gal4/UAS-shits	LC10 silencing	S3A
	LC10 / +	w−/+; +; R22D06-Gal4/+	Gal4 control for LC10 silencing	S3A
	T4T5 > shits	w−/+; R59E08-AD/UAS-shits; R42F06-DBD/UAS-shits	T4T5 silencing	S3B, G
	T4T5 / +	w−/+; R59E08-AD/+; R42F06-DBD/+	Gal4 control for T4T5 silencing	S3B, G
	T3 > shi	w−/+; +/UAS-shits; R29D11-Gal4/UAS-shits	T3 silencing	S3C
	T3/+	w−/+; +; R29D11-Gal4/+	Gal4 control for T3 silencing	S3C
	DNp35 > shits	w−/+; +/UAS-shits; R91C05-Gal4/UAS-shits	DNp35 silencing	S3E
	DNp35 / +	w−/+; +; R91C05-Gal4/+	Gal4 control for DNp35 silencing	S3E
	DNp09 > shits	w−/+; VT023490.p65ADZp/UAS-shits; R38F04.ZpGAL4DBD/ UAS-shits	DNp09 silencing	S3D
	DNp09 / +	w−/+; VT023490.p65ADZp/+; R38F 04.ZpGAL4DBD/+	Gal4 control for DNp09 silencing	S3D
	empty / shits (single)	w−/+; +/UAS-shits; pBDP-Gal4/UAS-shits	UAS control for synaptic silencing with single Gal4 drivers	S3A, C, E
Functional Imaging	LC11 > GCaMP6f	w−/+; R22H02-AD/UAS-GCaMP6f; R20G06-DBD/+	LC11 calcium imaging	3B–O, 5A, G, 6A, B, E–G, 7D, E
		w−/+; UAS-GCaMP6f/+; R22H02-Gal4/+		7F
	LC11 > ArcLight	w−/+; R22H02-AD/+; R20G06-DBD/UAS-ArcLight	LC11 voltage imaging	5A, F, 6, 7D
	LC11 > iGluSnFR	w−/+; R22H02-AD/+; R20G06-DBD/UAS-iGluSnFR	LC11 glutamate imaging	5A, E, 6A, B, E–G, 7D
	LC11 > iGABASnFR	w−/+; R22H02-AD/+; R20G06-DBD/UAS-iGABASnFR.F 102G	LC11 GABA imaging	5A, D
	LC11 > GACh3.0	+; UAS-GACh3.0; R22H02-Gal4	LC11 acetylcholine imaging	5A, C, 6A, B, D–G, 7D, F
	T3 > Chrimson, LC11 > GCaMP6f	norpA/Y; R22H02-LexA, lexAop2-GCaMP6f/+; UAS-Chrimson.mVenus/+	T3 optogenetic activation and LC11 calcium imaging	S5C
	T3 > GCaMP6f	w−/+; UAS-GCaMP6f/+; R29D11-Gal4/+	T3 calcium imaging	S5D–H
	T2 > jGCaMP7b	w−/+; +; R29D11-Gal4/UAS-jGCaMP7b	T2 calcium imaging	S6C
	T2>GCaMP6f	w−/+; UAS-LexA, lexAop2-GCaMP6f/+; R88C05-Gal4/+	T2 calcium imaging	S6D–G
Anatomical Imaging	T3-LC11 tGRASP	y,w−/+; R22H02-lexA/Cy0; R29D11-Gal4/UAS-tGRASPpre,lexAop2-tGRASPpost	Examination of synaptic connectivity from T3 to LC11 using targeted GFP reconstitution across synaptic partners (t-GRASP)	S5B
		y,w−/+; R22H02-lexA/s; R29D11-Gal4/UAS-tGRASPpre,lexAop2-tGRASPpost
	T2-LC11 tGRASP	y,w−/+; R22H02-lexA/Cy0; R88C05-Gal4/ UAS-tGRASPpre,lexAop2-tGRASPpost	Examination of synaptic connectivity from T2 to LC11 using targeted GFP reconstitution across synaptic partners (t-GRASP)	S6B
		y,w−/+; R22H02-lexA/s; R88C05-Gal4/UAS-tGRASPpre,lexAop2-tGRASPpost