PRMT5 imposed a dual repression on DDIT3 transcription to promote the malignancy of hepatocellular carcinoma

Table 1 Primers of cloning genes and real-time quantitative polymerase chain reaction

Primer	Sequence
PRMT5 qPCR	F: 5’-CGATCAGACCTACTGCTGTCA-3’
	R: 5’-CTCGGAGTTCCTGCGAATCT-3’
DDIT3 qPCR	F: 5’-CTCGCTCTCCAGATTCCAGTC-3’
	R: 5’-CTTCATGCGTTGCTTCCCA-3’
DDIT3 promoter	F: 5’-CCCTCGAGTCATTTTTAAAGAATGAGTTAAGGG-3’
	R: 5’-GATATCTGGGGAATGACCACTCTGTTTCC-3’
PRMT5 clone	F: 5’-CGGGATCCATGGCGGCGATGGCGGTC-3’
	R: 5’-CGACGCGTCGAGGCCAATGGTATATGAGCG-3’
DDIT3 clone	F: 5’-GGAATTCATGGAGCTTGTTCCAGCCACT-3’
	R: 5’-CGGGATCCTCATGCTTGGTGCAGATTCACCAT-3’

qPCR: Quantitative polymerase chain reaction; F: Forward primer; R: Reverse primer.

Table 2 Primers of short hairpin RNA targeted to PRMT5 and DDIT3

Primer	Sequence
shPRMT5-1	F: 5’-CCGGAGTACCAGCAGGCCATCTATACTCGAGTATAGATGGCCTGCTGGTACTTTTTTG-3’
	R: 5’-AATTCAAAAAAGTACCAGCAGGCCATCTATACTCGAGTATAGATGGCCTGCTGGTACT-3’
shPRMT5-2	F: 5’-CCGGGCCCAGTTTGAGATGCCTTATCTCGAGATAAGGCATCTCAAACTGGGCTTTTTG-3’
	R: 5’-AATTCAAAAAGCCCAGTTTGAGATGCCTTATCTCGAGATAAGGCATCTCAAACTGGGC-3’
shDDIT3-1	F: 5’-CCGGCTGCACCAAGCATGAACAATTCTCGAGAATTGTTCATGCTTGGTGCAGTTTTTG-3’
	R: 5’-AATTCAAAAACTGCACCAAGCATGAACAATTCTCGAGAATTGTTCA TGCTTGGTGCAG-3’
shDDIT3-2	F: 5’-CCGGTGAACGGCTCAAGCAGGAAATCTCGAGATTTCCTGCTTGAGCCGTTCATTTTTG-3’
	R: 5’-AATTCAAAAATGAACGGCTCAAGCAGGAAATCTCGAGATTTCCTGCTTGAGCCGTTCA-3’

shPRMT5: Short hairpin RNA of PRMT5; shDDIT3: Short hairpin RNA of DDIT3; F: Forward primer; R: Reverse primer.

Table 3 Chip primers of DDIT3 promoter

Primer	Sequence
ChIP primer 1	F: 5’-TTGCCAAACATTGCATCATC-3’
	R: 5’-AGCGCCTGGCTGTAATCTT-3’
ChIP primer 2	F: 5’-CAAACAGAATCGGGTCCACT-3’
	R: 5’-GGCAGAAGAATCGCTTGAAC-3’
ChIP primer 3	F: 5’-ACCACATGCCTCTGTTTTC-3’
	R: 5’-TGTGACTCCCTGTGTGAGGA-3’
ChIP primer 4	F: 5’-GTAGAGACGGGGTTTCACCA-3’
	R: 5’-TAGTCGGTCGTGAGCCTCTT-3’
ChIP primer 5	F: 5’-CCCTTCCAAAGCCACAGTCT-3’
	R: 5’-TTGTGACTGGAGTGGTGTGG-3’

ChIP: Chromatin immunoprecipitation; F: Forward primer; R: Reverse primer.