Knock-down of ACTL6A Promote Differentiation of NB4 Cells via the Notch1 Signaling Pathway

doi:10.13523/j.cb.20181201

China Biotechnology

2018, Vol. 38

Issue (12): 1-6 DOI: 10.13523/j.cb.20181201

Orginal Article

Knock-down of ACTL6A Promote Differentiation of NB4 Cells via the Notch1 Signaling Pathway

ZHONG Peng-qiang^1,²,LIU Bei-zhong^1,²(

),YAO Juan-juan^1,²,LIU Dong-dong²,YUAN Zhen²,LIU Jun-mei¹,CHEN Min^1,²,ZHONG Liang^2,^**(

)

1 Central Laboratory,Yongchuan Hospital of Chongqing Medical University,Chongqing 402160, China
2 Key Laboratory of Chongqing,Key Laboratory of Medical Diagnostics of Ministry of Education,Department of Laboratory Medicine,Chongqing Medical University,Chongqing 400016,China

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Abstract

To explore the role of ACTL6A in NB4 cell differentiation and its related mechanisms.When we used ATRA to induce differentiation of NB4,the expression levels of ACTL6A and CD11b were detected by Western blotting.Morphological changes of NB4 cells were observed by Wright staining.The positive rate of CD11b was detected by flow cytometry.Immunofluorescence was used to detect location of ACTL6A in NB4 cells.The results showed that the protein expression of CD11b increased after ACTL6A was knocked down.Differentiation changes were observed by Wright staining.Immunofluorescence detection showed that ACTL6A was mainly distributed in the nucleus.The expression levels of Notch1,Hes1 and Sox2 proteins were significantly down-regulated,analysed by Western blotting. Therefore,the knockdown of ACTL6A can promote the differentiation of NB4 cells.The mechanism is involved in the inhibition of Notch1 signaling pathway.

Key words： ACTL6A NB4 cells Differentiation Notch1 signal pathway

Received: 30 July 2018 Published: 10 January 2019

ZTFLH:

Q813

Corresponding Authors: Liang ZHONG E-mail: Liubeizhong@cqmu.edu.cn

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	Peng-qiang ZHONG
	Bei-zhong LIU
	Juan-juan YAO
	Dong-dong LIU
	Zhen YUAN
	Jun-mei LIU
	Min CHEN
	Liang ZHONG

Cite this article:

ZHONG Peng-qiang,LIU Bei-zhong,YAO Juan-juan,LIU Dong-dong,YUAN Zhen,LIU Jun-mei,CHEN Min,ZHONG Liang. Knock-down of ACTL6A Promote Differentiation of NB4 Cells via the Notch1 Signaling Pathway. China Biotechnology, 2018, 38(12): 1-6.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20181201 OR https://manu60.magtech.com.cn/biotech/Y2018/V38/I12/1

Fig.1 The expression levels of ACTL6A during ATRA-induced granulocytic differentiation of NB4 Note:Western blotting analysis the protein expression levels of ACTL6A and CD11b in NB4 and HL-60 cells treated with ATRA(1 μmol/L)for 72hours.

Fig.2 The knockdown of ACTL6A promotes granulocytic differentiation of NB4 (a) The mRNA expression level of ACTL6A were detected by qRT-PCR (b) Morphological observation of shACTL6A cells analysis by Wright staining experience

Fig.3 The knock down of ACTL6A promote differentiation of NB4 induced by ATRA Note: Flow cytometry was used to detect the level of CD11b positive rate in NB4 cells

Fig.4 Immunofluorescence analysis (×400) of endogenous ACTL6A (red) localizations in NB4 cells after treated with ATRA for 72hours

Fig.5 Western blotting was used to detect the level of ACTL6A and CD11b

Fig.6 Western blotting was used to detect the level of ACTL6A,CD11b,Notch1 and Hes1


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