TABLE II.

Oligonucleotides Used in this Study

Name of DNA	Oligonucleotides	Restriction sites
For constructs
a) Nt2NKCC2(51–179)/pGilda	forward: gg GAA TTC ggg gat gaa gct cag aaa ag	EcoRIb
	reverse: cc CTC GAG cca tcc aaa ctt cac aac acc	XhoI
b) Ct1NKCC2(657–836)/pGilda	forward: cc GAA TTC gct ctt tcc tac gtg agt gc	EcoRIb
	reverse: tg CTC GAG cct gtt cta atc tct cta att cc	XhoI
d) Ct1NKCC2(671–836)/pGilda	forward: c aat gct ctg GAA TTC acc aca gtg gaa gac	EcoRIc
	reverse: gtc ttc cac tgt ggt GAA TTC cag agc att g	EcoRI
e) Ct1NKCC1(759–947)/pGilda	forward: tc acC CCG GGa tcc tct aca caa gca	XmaIb
	reverse: ca tgC CAT GGt gcc agg aga ttt cta	NcoI
f) Ct1NKCC1(834–926)/pGilda	forward: ca GAA TTC atg tcc atc gat caa gcca	EcoRIb
	reverse: c ACT CGA gtc cag acc ttc ttt tag gcga	BsRI
h) Ct2NKCC2(830–1099)/B42AD	forward: gg GAA TTC gaa tta gag aga tta gaa cag gag	EcoRIb
	reverse: gg CTC GAG agt taa aat gta ttc caa tct ttc	XhoI
i) Ct2NKCC2(910–1058)/B42AD	forward: c att gtc ctg agc ctt CCC GGG caa gaa agg g	XmaIc
	reverse: c cct ttc ttg CCC GGG aag gct cag gac aat c	XmaI
For sequencing
j) pGilda	forward: cgt cag cag agc ttc acc att ga
k) pB42AD	forward: cc agc ctc ttg ctg agt gga gat ga
	reverse: gca aag tag aca agc cga caa cc
For expression studies
l) “A” exon (RT-PCR)	forward: cg gga att ggt ctt gga g
	reverse: cc tcc acg aac aaa ccc g
m) “F” exon (RT-PCR)	forward: cg gga att ggt ctg ggc g
	reverse: cc tcc tcg cac cac tcc g
n) “AF” exon (RT-PCR)	forward: cg gga att ggt ctt gga g
	reverse: cc tcc tcg cac cac tcc g
o) “A” oligoprobe (in situ)	forward: tt ctt ctt tcc acc atg gta acc tct at
	reverse: at aga ggt tac cat ggt gga aag aag aa
p) “F” oligoprobe (in situ)	forward: tt ggc ctg agc gta gtt gtg aca aca ct
	reverse: ag tgt tgt cac aac tac gct cag gcc aa

They are all written 5′ to 3′. Added restriction sites are designated by capital letters. Note that Ct1_{NKCC2(657–782)}/pGilda is not shown in the list because it was produced by deleting a portion of Ct1_{NKCC2(657–836)} between BplI, a naturally occurring restriction site in huNKCC2 and XhoI in the pGidla cloning site.

^aPrimers were available from a previous study (Simard et al., 2004a).

^bPrimers were used for RT-PCR.

^cPrimers were used to introduce substitutions with the Quick Change Mutagenesis Kit (Stratagene).