Midline Molecular Data

Stage 17 Gene Expression Table


	AMG	MP1s	H-cell	H-cell sib	mVUMs	iVUMs	MNB	MNB prog
	AA41-lacZ	5-HT1A	5-HT1A	5-HT1A	807-Gal4 (Sub)	Cas (Sub)	ase	Cas (Sub)
	argos	C544-Gal4	CG33528	Elav	Cas (Sub)	CG15236	En	CG8394
	Runt	CG6201	DAT	fkh	CG33528	CG8394	Sim	Elav (Sub)
	Sim	Elav	Elav	Glu-RI	Elav	Elav	wor	En
	slit-Gal4	fkh	Glu-RI	per-Gal4	Futsch	En		fkh
	sna	Hb	NPFR1	Sim	Glu-RI	fkh		Gad1
	SoxN	HGTX	ple	VGlut	MzVUM-Gal4	Gad1		per-Gal4
	vvl	odd	TH-Gal4	X55-lacZ	SNF4Ag	per-Gal4		Pros-nuc (Sub)
	wrapper	Pdf	SNF4ag		Tbh	Sim		Sim
	X55-lacZ	Runt	SoxN		Tkr (Sub)	Tkr (Sub)		Spdo
		X55-lacZ	X55-lacZ		VGlut	X55-lacZ		Tkr (Sub)
					X55-lacZ			X55-lacZ
					zfh1

	All Midline Cells
	SimGal4>UAS-tau-lacZ

	Not Expressed
	Cad74A	Dichaete	fd59a	nub	pxb	Spdo
	CG32244	esg	hh	pdm2	Slp1	Vnd
	CG8965	Eve	l(1)sc	Prd	slp2	wg
___________________________________________________________________________________
	MG – midline glia. sub - indicates that a marker is expressed in a subset of cells within this group. Pros-nuc indicates that Pros protein is observed in the nucleus, while Pros-cort indicates that Pros protein is observed in a cortical crescent. Italics indicate that an RNA probe was used for in situ hybridization to detect RNA, while normal text indicates that an antibody was used to detect protein by immunostaining. lacZ and Gal4 line (crossed to UAS-lacZ) expression was visualized by staining with an antibody to ?-galactosidase. Not expressed in midline cells – refers to genes used in this study that are not expressed in midline cells at this stage. (#) - We occasionally observe per-Gal4 in one ventral en+ MNB progeny. * - X55-lacZ is expressed at low levels in MG.

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Stage 13 Gene Expression Table


	AMG	PMG	MP1s	H-cell	H-cell sib	mVUMs	iVUMs	MNB	MNB prog
	argos	argos	C544-Gal4	CG33528	Elav	Cas (Sub)	Cas (Sub)	ase	ase (Sub)
	AA41-lacZ	Cad74A	CG6201	Elav	fkh	CG33528	CG15236	En	Cas (Sub)
	CG31145	CG31145	Elav	nub	nub	Elav	Elav	Pros-cort	Elav (Sub)
	CG32244	CG32244	fkh	pdm2	pdm2	fd59a	En	Spdo	En
	Dichaete	Dichaete	Hb	ple	Sim	Futsch	fkh	Sim	fd59a (Sub)
	Runt	En	HGTX	SNF4ag	X55-lacZ	MzVUM-Gal4	per-Gal4	wor	Pros-nuc
	Sim	Sim	nub	SoxN		SNF4Ag	Sim		Sim
	slit-Gal4	slit-Gal4	odd	X55-lacZ		Tkr (Sub)	Tkr (Sub)		Spdo
	sna	sna	pdm2			X55-lacZ	X55-lacZ		Tkr (Sub)
	SoxN	SoxN	Runt						wor (Sub)
	vvl	vvl	X55-lacZ						X55-lacZ
	wrapper	wrapper
	X55-lacZ	X55-lacZ

	All Midline Cells
	SimGal4>UAS-tau-lacZ

	Not Expressed
	5-HT1A	DAT	Gad1	l(1)sc	Prd	slp2	VGlut
	807-Gal4	esg	Glu-RI	NPFR1	pxb	Tbh	Vnd
	CG8394	Eve	hh	Pdf	Slp1	TH-Gal4	wg
___________________________________________________________________________________
	# - The AA41-lacZ enhancer trap line has an insertion in the CG31145 gene. In situ hybridization with a CG31145 RNA probe confirms that it is expressed at stage 13 in AMG and PMG. * - X55-lacZ is expressed at low levels in AMG and PMG.

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Stage 11 Gene Expression Table


	AMG	PMG	MP1s	MP3s	VUMs	MNB
	argos	argos	Elav	Elav	ase	ase
	Cad74A	Cad74A	Hb	nub	Cas	Cad74A
	CG32244	Cas	nub	pdm2	Elav (Sub)	Cas
	Runt	CG32244	odd	Pros-nuc	En	En
	sna	En	pdm2	SoxN (#)	Pros-nuc	slp2
	SoxN	sna	Runt	Spdo (trans)	Spdo (trans)	wg
	vvl	SoxN	Pros-nuc	wor (trans)	Tkr (trans)	l(1)sc
	wrapper	Tkr	Spdo (trans)	zfh1 (trans)	wor (trans)	Pros-cort (Trans)
		vvl	wor (trans)		zfh1 (trans)	Spdo
		wrapper	zfh1 (trans)			Tkr
						wor
						zfh1 (trans)
	All Midline Cells
	Dichaete	HGTX	SimGal4>UAS-tau-lacZ
	esg	Sim	X55-lacZ

	Not Expressed
	5-HT1A	CG8965	fkh*	NPFR1	slit-Gal4	VGlut
	807-Gal4	CG15236	Futsch	Pdf	Slp1	Vnd
	AA41-lacZ	CG33528	Gad1	per-Gal4	slp2	wg
	C544-Gal4	DAT	Glu-RI	ple	SNF4a
	CG6201	Eve	hh	Prd	Tbh
	CG8394	fd59A*	MzVUM-Gal4	pxb	TH-Gal4
________________________________________________________________________________________
	(trans) indicates that a gene is transiently expressed in this cell type. * - fd59A and fkh initiate expression at late stage 11/early s12 in a subset of midline cells. (#) – SoxN may only be expressed in a single cell at mid-stage 11. If so, this suggests that H-cell and H-cell sib are beginning to acquire their respective fates at this stage.

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Stage 9 Gene Expression Table


	Row A	Row B	Row C	Row D	Row E	Row F	Row G	Row H
	l(1)sc	l(1)sc	pxb	pxb	l(1)sc	l(1)sc	En	En
	odd	odd	Runt	Runt	Prd	Prd	Eve*	hh
				Slp1	pxb	pxb	hh
				slp2	Runt	Slp1
					Slp1	slp2
					slp2	wg
					wg
	All Midline Cells
	Cas	HGTX	Vnd
	Dichaete	Sim	vvl

	Not Expressed
	5-HT1A	Cad74A	CG33528	Futsch	nub	sim-Gal4	Tbh	X55-lacZ
	807-Gal4	CG6201	DAT	Gad1	Pdf	slit-Gal4	TH-Gal4	zfh1
	AA41-lacZ	CG8394	Elav	Glu-RI	pdm2	sna	Tkr
	argos	CG8965	esg	Hb	per-Gal4	SNF4a	VGlut
	ase	CG15236	fd59A	MzVUM-Gal4	ple	SoxN	wor
	C544-Gal4	CG32244	fkh	NPFR1	Pros	Spdo	wrapper
_______________________________________________________________________________________________________________________
	In this table, midline cells have been divided in 8 rows (A-H) of 2 cells/row along the A-P axis. However, midline cells often do not line up in pairs and there is also variability in the A-P position. * - indicates that Eve is expressed in every other segment at this stage.

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