Tonicity inversely modulates lipocalin-2 (Lcn2/24p3/NGAL) receptor (SLC22A17) and Lcn2 expression via Wnt/β-catenin signaling in renal inner medullary collecting duct cells: implications for cell fate and bacterial infection

Table 2 Protocols for reverse transcription semi-quantitative PCR

	Gapdh	Ctnnb1 (β-catenin)	Lcn2	Lcn2-R (Slc22a17/24p3-R)	hLCN2-R isoform B
Accession number	NM_001289726.1	NM_007614.3	NM_008491.1	NM_021551.4	(NM_016609.4)
Forward primer (5‘→3‘)	AGGGCTCATGACCACAGT	TTAAACTCCTGCACCCACCAT	CCACCACGGACTACAACCAG	CAGCCACCTCCTAACCGCTGTG	GCCATTCGCCACTGCTAC
Reverse primer (5‘→3‘)	TGCAGGGATGATGTTCTG	AGGGCAAGGTTTCGAATCAA	AGCTCCTTGGTTCTTCCATACA	CTCCCACTAGGCTCAAAGGCTGCT	GGAGAAGAGCCCAAGGACAGA
Activation	5 min 95^oC	5 min 95^oC	5 min 95^oC	5 min 95^oC	5 min 95^oC
Cycle number	22	27	30-32	27-28	22
Denaturation	30 sec 94^oC	30 sec 94^oC	30 sec 94^oC	30 sec 94^oC	30 sec 94°C
Annealing	30 sec 60^oC	30 sec 60^oC	30 sec 60^oC	30 sec 60^oC	30 sec 55°C
Extension	30 sec 72^oC	30 sec 72^oC	30 sec 72^oC	30 sec 72^oC	30 sec 72°C
Final Extension	7 min 72^oC	7 min 72^oC	7 min 72^oC	7 min 72^oC	10 min 72°C
PCR product (bp)	112	73	100	86	255

ISSN: 1478-811X