Lipopolysaccharide triggers nuclear import of Lpcat1 to regulate inducible gene expression in lung epithelia

Table 1 Primers used in quantitative polymerase chain reaction

Gene symbol	Forward primer	Reverse primer
L19	AGAGCTTGGAGTGCACAGGT	GGGACAGTCCAAACTGGGTA
TGF-b1	GACTCTCCACCTGCAAGACC	GACTGGCGAGCCTTAGTTTG
IL-13Ra1	CACAGAAGTTCAGCCACCTG	AATTTGGACTGGCTCCTTCA
ILRN	GTTGGAAGGCAGTGGAAGAC	GCATCTTGCAGGGTCTTTTC
IL4	CCAAGGTGCTTCGCATATTT	ATCGAAAAGCCCGAAAGAGT
SPHK1	GCTGTGAGGCTGGTGTTATG	ATATGCTTGCCCTTCTGCAT
EDG7	ATTGCCTCTGCAACATCTCG	ATGAAGAAGGCCAGGAGGTT
IL-13Ra2	TTGGTCTGCTCTTGGAAACC	CAGCACACTGTAAGGCATGA
Autotaxin	GAAACAGCACCTTCCCAAAC	AAGGTTTCCTTGCAACATGC
LPA4	ACTGCGTTCCTCACCAACAT	CGATCGGAAGGGATAGACAA
sST2	CATGGCATGATAAGGCACAC	GATGCAGTGCACAGCTGATT
LPCAT1	CGACTGAGCGCCCTGCAGAA	AAGGGCCAGGCCAGCAGCAT
ST2L	GGAACGATGGCAAGCTCTAC	GGCAGAGTGTGGTGAACAAA
EDG1	CTGTTAGATGTGGGCTGCAA	ATGATGGGGTTGGTACCTGA
MMP9	CTTCGAGGGACGCTCCTATT	CAAATTTGCCGTCCTTATCG
IL-1β	CAGGCAGGCAGTATCACTCA	AGGTGCTCATGTCCTCATCC
IL33	CCTTCTCGCTGATTTCCAAG	GCTGAACAGAACGTGTGCAT
IL-13	CAGCAGCTTGAGCACATTTC	ATAGGCAGCAAACCATGTCC
EDG8	ACACCAAATGCCCAGCTTAC	TGGAGCACTGTGCAAAAGTC
EDG2	TCAACCTGGTGACCTTTGTG	GGTCCAGAACTATGCCGAGA
IL4R	TCACAGAGCAGCCTTCACAC	TTGTCTGCAAGGACAAGTGG
LPA5	GCTCCAGTGCCCTGACTATC	CAGAGCGTTGAGAGGGAGAC
CCTa	GGAAACTCCGGGTAGTGGGTGC	TTCCCTAGCTGTCACTGGCCCA
COX2	CCCCCACAGTCAAAGACACT	GGCACCAGACCAAAGACTTC
CTTN	AGCAGAGATGGGGTGCTAAA	TTGAGCGTCTGGTGTTCTTG
SGPP2	CATCTCCTTCACCCTCCTCA	CCAGCGTAGAGAACACCACA
SGPL1	AACTCTGCCTGCTCAGGGTA	CTCCTGAGGCTTTCCCTTCT
IL-6	CCGGAGAGGAGACTTCACAG	TCCACGATTTCCCAGAGAAC
MET	CAGCCATCCCAATGTTCTCT	AATTTCGCAGATCTCCATGC
EDG3	TGCCTTATGAACCCTGGAAG	AAGAGCAAATGCCATCAGGT
EDG6	CTCCAAGGGCTATGTGCTCT	ATTGGCTCGGACCACTCTAA
CADHERIN	GCACTCTTCTCCTGGTCCTG	TATGAGGCTGTGGGTTCCTC
SGPP1	GAGCAACTTGCCCGCTCTACT	AAGGGGTCGAGATTCCAGAT
SPHK2	ACTGCTCGCTTCTTCTCTGC	GCCACTGACAGGAAGGAAAA

The sequences of all the primers are from 5’- to -3’.

Table 2 Lpcat1 regulates gene expressionreaction

Gene symbol	Vector	Lpcat1 shRNA	Lpcat1 overexpression
IL-1β	2.29	119.70	-29.79
IL1RN	-1344.87	8.22	-8579.43
IL4	-3029.24	-2.65	1.86
IL4R	5.79	1.17	-7.18
IL6	7.38	10.88	998.30
IL13	-19.56	-4.02	1237.60
IL13Ra1	-171.65	65.34	-544.96
IL13Ra2	-2845.00	53.32	207.94
IL33	20.48	1.38	-1.46
Autotaxin	17.47	4.17	-3.50
sST2	-4.11	1667.34	21.91
ST2L	2.42	138.14	781.44
TGF-b1	-1.07	-6.35	-14.69
COX2	13.33	2.40	8.0
MMP2	-13.55	443.67	-565.48
MMP9	7.00	124.79	-1.99
EDG1	-93.05	541.37	-1231.90
EDG2	-6.85	-1.26	-48.50
EDG3	2.07	30.06	-3.70
EDG4	-1.27	83.48	-36.25
EDG5	-1.59	50.21	-198.09
EDG6	-70.20	-1.56	-343.30
EDG7	-180.60	-1978.24	2189.93
EDG8	-441.62	-2.52	19.79
CCT	-17.71	-227.54	6397.68
CCTN	-1.49	1.10	-22.63
SPL1	5.62	7.11	186.54
SGPP1	-2.80	-2.30	-64.73
SGPP2	-26.85	67.18	305.14
SGPL1	-1.56	-1.17	-65.04
SPHK1	-8.28	22.37	-133.74
SPHK2	1.59	-2.63	1223.39
LPA4	-21.96	15.63	-110.15
LPA5	1.15	275.01	3590.58
MET	1.72	-8.00	-38.49
E-CADHERIN	-2.62	-18.77	-155.41
L19	- 667.15	6.09	-2752.77
LPCAT1	3.04	-5.24	132.21

Gene expression fold changes were adjusted by baseline expression of each gene in the untreated normal cells.