NBPF1, a tumor suppressor candidate in neuroblastoma, exerts growth inhibitory effects by inducing a G1 cell cycle arrest

Table 2 Overview of shRNAs and siRNAs used in this study with the aim to silence the expression level of NBPF family members

Knockdown type	#	Target seq (5' to 3')	Targets only NBPF1		Exon type
Knockdown type	#	Target seq (5' to 3')	No	Yes	Exon type
shRNA_19-mer	1	CCUCUUCUGCCACAAACGUUU	X		1
	2	GGAAUGUGCCAUCACUUGU	X		5
	3	GGAAGUCCCUGAGGACUCA	X		6
	4	GAAGAAGAUCAACGAAGAA		X	12
	5	CUUACUUUGAUGGGAACAA		X	14
shRNA_29-mer	6	UACCUCUUCUGCCACAAACGUCAGCAUGGU	X		1
	7	AUGUCAGGAUGCUGUAAACAUUCUCCCAG		X	5
	8	AACAUCAACAUCACCUUUGAGGAAGACAA		X	5
	9	UGACUCCAACCAGCCUCACAAGAACAUCA	X		5
	10	GGAGAAGACGGUGUACCAGCCUCACAAGA	X		5
	11	ACCAGUCUUACAGCGGCACAUUUCACUCA		X	8
	12	CCUGACUCAUUCCAGCACUACAGAAGUGU	X		14
	13	AGCUGGCAGAGAACAAACAGCAGUUCAGA	X		1
	14	AUGCUGAGGAAUGAGCGACAGUUCAAGGA	X		2
	15	UAAGGGAGAAGUUACGGGAAGGGAGAGAU	X		3
	16	CUUCCUGGCCAACCAGCAGAACAAAUACA	X		1
siRNA pool	17	GAAUCGUACUGCUAAGAAU		X	5' UTR
		ACAGAGACAGACAAAGAUA	X		5' UTR
		GAUGGCCAAUCUUUCCUAA	X		5' UTR
		GCAAGUCCAGGUCAUACUG	X		5' UTR

Given are the target sequence, the predicted specificity (NBPF1-specific or broad NBPF specificity) and location in exons (for exon type, see ref. [4]) or 5’UTR. shRNAs 1 to 5 represent shRNAs with a stem length of 19 bps, whereas shRNAs 6 to 16 represent shRNAs with a stem length of 29 bps. For shRNAs 1 to 8, the expression cassettes of the shRNAs were created by us by a double PCR-approach, whereas shRNAs 9 to 16 are HuSH retroviral silencing vectors from Origene (Origene Technologies, Rockville, MD, USA). siRNA pool 17 represents an On-Target plus siRNA pool from Dharmacon (Thermo Fisher Scientific, Lafayette, CO, USA)

ISSN: 1471-2407