Diet high in fructose leads to an overexpression of lipocalin-2 in rat fatty liver

Table 1 List of primers that were used in this study

Name	Forward 5’-3’	Reverse 5’-3’
Lcn-2	GGAATATTCACAGCTACCCTC	TTGTTATCCTTGAGGCCCAG
Il-8	GTGTCCCCAAGTAATGGAGAA	CGCCTACCATCTTTAAACTGC
α2-m	CTGTCACTCATCCTGTTGTC	ATCTCCTTCTTCGTGTCCTG
Glut5	TGCAGAGCAACGATGGAGAAA	ACAGCAGCGTCAGGGTGAAG
Tnf-α	AAATGGGCTCCCTCTCATCAGTTC	TCTGCTTGGTGGTTTGCTACGAC
Mcp-1	CTCACCTGCTGCTACTCATTCACT	TTCCTTATTGGGGTCAGCAC
Lep-r	GTTCTGGCCATCAATTCCAT	GCCCTCTGGTTGCTTTGTAT
Actb	ACCACCATGTACCCAGGCATT	CCACACAGAGTACTTGCGCTCA
Ubc	CACCAAGAAGGTCAAACAGGAA	AAGACACCTCCCCATCAAACC

Table 2 List of antibodies used in this study

Name	Clone	Species	Company	Dilution	Use
HNE	PC	Rabbit	Abcam	1:100	WB
LCN-2	MC	Mouse	Novus biological	1:250	WB
	PC	Goat	R&D	1:100	IF
GRP-78	PC	Rabbit	Novus Biocompare	1:800	WB
CD14	PC	Rabbit	Antibodies-online	1:700	WB
Casp 9	PC	Rabbit	Chemicon international	1:1200	WB
Cyt c	MC	Mouse	Millipore	1:600	WB
PGC-1α		Goat	Abcam	1:200	WB
IκB1α	MC	Rabbit	Abcam	1:10000	WB
β-actin	MC	Mouse	Sigma Aldrich	1:5000	WB
MPO	PC	Rabbit	Dako, A0398	1:100	IF
ED1	MC	Mouse	AbD Serotec	1:100	IF
Anti-rabbit-HRP	PC	Swine	Dako	1:1500	WB
Anti-mouse-HRP	PC	Rabbit	Dako	1:1500	WB
AlexaFluor-555-conjugated anti-goat IgG		Donkey	Invitrogen	1:500	IF
AlexaFluor-488-conjugated anti-mouse IgG		Donkey	Invitrogen	1:500	IF
AlexaFluor-488-conjugated anti-rabbit IgG		Donkey	Invitrogen	1:500	IF

WB: Western blotting; MC: Monoclonal; PC: Polyclonal; IF: Immunofluorescence stain.

Table 3 Data of plasma biochemistry of Sprague-Dawley rats within 8 wk of feeding

Parameter	Time point/ Group (wk)	Co	LDC	L-HFr
Glucose (mg/dL)	4	152 ± 12	182 ± 5	243 ± 8b
	8	210 ± 7	244 ± 5	292 ± 7b
TG (mg/dL)	4	26 ± 5	74 ± 8a	78 ± 13a
	8	44 ± 13	62 ± 15	127 ± 21a
Cholesterol (mg/dL)	4	55 ± 2	58 ± 4	58 ± 3
	8	47 ± 4	52 ± 4	53 ± 5
HDL-C (mg/dL)	4	49 ± 1	43 ± 2	46 ± 1
	8	44 ± 1	45 ± 3	32 ± 2d
Uric acid (mg/dL)	4	0.6 ± 0.0	0.9 ± 0.2	0.5 ± 0.0
	8	1.1 ± 0.0	0.5 ± 0.1	2.2 ± 0.3b
ALT (U/L)	4	41 ± 8	43 ± 3	69 ± 7a
	8	55 ± 8	48 ± 4	80 ± 8a
AST (U/L)	4	67 ± 5	74 ± 2	136 ± 8b
	8	90 ± 7	117 ± 13	166 ± 10b
Leptin (ng/mL)	4	1.8 ± 0.2	3.0 ± 0.3	5.0 ± 0.9b
	8	3.7 ± 0.9	5.7 ± 2.1	12.1 ± 1.4b

All animals were overnight fasted just before sacrificing.

^aP < 0.05,

^bP < 0.01 compared to controls. Data are expressed as mean ± SEM of 4 rats per group. Co: Chow (control); LDC: Lieber-DeCarli liquid diet; L-HFr: LDC + 70% cal fructose; ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; TG: Triglycerides; LDL-C: Low density lipoprotein cholesterol.