Caffeic acid phenethyl ester up-regulates antioxidant levels in hepatic stellate cell line T6 via an Nrf2-mediated mitogen activated protein kinases pathway

Table 1 Primers used for real-time PCR analysis

Gene	Primer sequence	Tm (°C)	Size (bp)	Source
Nrf2
F	5’-CTGCCATTAGTCAGTCGCTCTC-3’	57.3	22	Rat
R	5’-TCGGCTGGGACTTGTGTTC-3’	57.7	19	Rat
SOD1
F	5’-GCTTCTGTCGTCTCCTTGCT-3’	55.7	20	Rat
R	5’-CTCGAAGTGAATGA CGCCCT-3’	55.8	20	Rat
CAT
F	5’-TGGCTATGGCTCACACACCTTC-3’	59.3	22	Rat
R	5’-GAGGCCA TAATCCGGGTCTTC-3’	57.7	21	Rat
GSTs
F	5’-GTGGAGATTGACGGGATGAA -3’	54.2	20	Rat
R	5’-CGGTCTTGGCTTCTCTTTGG -3’	56.0	20	Rat
β-actin
F	5’-GGAGATTACTGCCCTGGCTCCTA-3’	60.2	23	Rat
R	5’-GACTCATCGTACTCCTGCTTGCTG-3’	59.3	24	Rat

Table 2 Antibodies used for Western blot analysis

Antibody (dilution)	Clone	Species	Specificity	Source
Nrf2	M	Rabbit	Immunogen: ag9489	ProteinTech, United States
(16396-1-AP 1:500)
P38MAPK	M	Rabbit	Synthetic peptide corresponding to residues near the C-terminus of human p38 MAP kinase was used as immunogen. Predicted to react with CSBP1 splice form based on sequence homology	EPITOMICS, United States
(ab32142, 1:500)
P-p38MAPK	M	Rabbit	Phospho-P38 MAPKinase (Thr180/Tyr182) (3D7) rabbit mAb detects endogenous levels of p38MAP kinase only when dually phosphorylated at Thr180 and Tyr182. This antibody does not cross-react with the phosphorylated forms of either p42/44 MAPK or SAPK/JNK	Cell Signaling, United States
(#4511, 1:500)
Erk1/2	M	Rabbit	P44/42 MAP kinase (137F5) rabbit mAb detects endogenous level of total P44/42 MAP kinase (Erk1/Erk2) protein. The antibody does not cross-react with JNK/SAPK or p38 MAP kinase	Cell Signaling, United States
(#4695, 1:500)
Erk1phospho(pY204)/Erk2phospho	M	Rabbit	A phospho-specific peptide corresponding to residues surrounding tyrosine 204 of human Erk1. This antibody detects Erk1 phosphorylated at tyrosine 204	EPITOMICS, United States
(pY187)
(ab76299 1:500)
SAPK/JNK	M	Rabbit	SAPK/JNK(56G8) rabbit mAb detects endogenous levels of total SAPK/JNK protein	Cell Signaling, United States
(#9258 1:500)
JNK1phospho(pT183) /JNK2phospho(pT221) (ab124956 1:500)	M	Rabbit	A phospho-specific peptide corresponding to residues surrounding threonine 221 of human JNK3 was used as immunogen. The antibody detects JNK1 (pT183), JNK2 (pT183) and JNK3 (pT221)	EPITOMICS, United States
α-SMA	M	Rabbit	A synthetic peptide corresponding to N-terminus of human actin was used as immunogen	EPITOMICS, United States
(ab124964,1: 1000)
Collegen-1	P	Rabbit	KLH conjugated synthetic peptide derived from human Collagen I C-terminal propeptide	Bioss, China
(bs-0578R 1:500)
β-actin	M	Mouse	β-actin is recommended for detection of β-actin of mouse, rat, human, chicken, dog, pig, rabbit	SANTA CRUZ, United States
(sc-47778,
1:1000)

M: Monoclone; P: Polyclone.