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Fig. 5 | BMC Cancer

Fig. 5

From: The autophagy GABARAPL1 gene is epigenetically regulated in breast cancer models

Fig. 5

GABARAPL1 expression I controlled by CREB-1. a Scheme describing the position of primers used (in regard of the putative initial transcription site (+1)) and putative CRE (CREB-1 response elements) sites in the GABARAPL1 promoter. b and c Luciferase activity measured using a Luciferase assay System Kit in MCF-7 cells transfected with empty pGL3 plasmid,−336/+241-GABARAPL1-promoter-pGL3 plasmid,–659/+241-GABARAPL1-promoter-pGL3 plasmid, pCDNA3.1-CREB-1 or treated with (10 μM) forskolin. c Bottom : expression of CREB-1 using IF in cells transfected or not with the pCDN3A.1-CREB-1 vector. d Recruitment of CREB-1 on–659/+241-GABARAPL1-promoter-pGL3 plasmid using ChIP experiment and anti-CREB-1 antibody in MCF-7 cells transfected with–659/+241-GABARAPL1-promoter-pGL3 and pCDNA3.1-CREB-1 plasmids (I: input; IgG : negative control of IP). e Half-life of GABARAPL1 mRNA using qRT-PCR following Actinomycin D treatment in MCF-7 cells. f Effects of CREB-1 overexpression (following pCDNA3.1-CREB-1 plasmid transfection) and/or 5-aza-CdR /TSA treatment on GABARAPL1 expression using qRT-PCR in MCF-7 cells. g Effects of CREB-1 overexpression (following pCDNA3.1-CREB-1 plasmid transfection) and/or 5-aza-CdR/TSA treatment on CREB-1 recruitment in GABARAPL1 promoter using ChIP experiment and an anti-CREB-1 antibody (I: input; IgG : negative control). Differences were quantified using a t-tests. GL1: GABARAPL1

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