Inducible Expression of UEV1A Mutant

The human UEV1A open reading frame (ORF) was amplified and cloned into a Dox-inducible Tet-ON plasmid pcDNA4.0/TO/HA(+) as described previously [20 (link)]. The mutated Ubc13-binding site (F38E) in Uev1A was designed based on a previous study with Mms2-F13E [7 (link)]. FOXO1 (MYC-DDK-tagged)-human forehead box O1 (NM_002015) plasmid (RC200477) and pCMV6-Entry vector (PS100001) were from Origene. The human BIM ORF was PCR-amplified as a KpnI-XhoI fragment and cloned into plasmid pcDNA4.0/TO/HA(+). The modified sequence for UEV1 shRNA (sc-38606-v) and negative control shRNA (sc-108080) delivered by lentiviral particles were from Santa Cruz Biotechnology, Inc. The lentiviral particle infection of colon cancer cells was performed following instructions of the supplier.

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Wu Z., Niu T, & Xiao W. (2019). Uev1A promotes breast cancer cell survival and chemoresistance through the AKT-FOXO1-BIM pathway. Cancer Cell International, 19, 331.

Publication 2019

pCMV6-Entry vector (PS100001) (sc-108080)

Binding site Cancer of the colon Cells Forehead Human Infection Plasmid Shrna Ubc13 Vector

Corresponding Organization :

Other organizations : University of Saskatchewan

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Variable analysis

independent variables

Uev1A open reading frame (ORF)
Mutated Ubc13-binding site (F38E) in Uev1A
UEV1A shRNA
Negative control shRNA

dependent variables

Not explicitly mentioned

control variables

Dox-inducible Tet-ON plasmid pcDNA4.0/TO/HA(+)
FOXO1 (MYC-DDK-tagged)-human forehead box O1 (NM_002015) plasmid (RC200477)
PCMV6-Entry vector (PS100001)
Human BIM ORF cloned into pcDNA4.0/TO/HA(+)

controls

Negative control shRNA (sc-108080)

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