Stable Knockdown of p53 and AMPK in CRC

As described in our previous studies [29 (link)], lentiviral particles containing the p53-shRNA-1 (Santa Cruz, sc-29435-V), p53 shRNA-2 (Genechem, Shanghai, China) or AMPKα1 shRNA (Santa Cruz, sc-29673-V) (10 μL/well) were added to cultured CRC cells for 24 h. Afterwards, cell culture medium was replaced with puromycin (0.5 μg/mL)-containing complete medium. The medium was renewed every 2-3 days until resistant stable colonies were formed. Expression of targeted protein (AMPKα1 and p53) was always determined by Western-blots in resulting stable cells. Same amount of scramble control shRNA lentiviral particles (Santa Cruz, sc-108080) were added to control cells.

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Lu P.H., Chen M.B., Ji C., Li W.T., Wei M.X, & Wu M.H. (2016). Aqueous Oldenlandia diffusa extracts inhibits colorectal cancer cells via activating AMP-activated protein kinase signalings. Oncotarget, 7(29), 45889-45900.

Publication 2016

sc-108080

Cell culture Cells Cultured cells Puromycin Sc 10 Shrna Western blots

Corresponding Organization :

Other organizations : Nanjing Medical University, Nanjing University of Chinese Medicine, Wuxi People's Hospital, Jiangsu University, First People's Hospital of Kunshan, First Affiliated Hospital of Fujian Medical University, Fujian Medical University, Jiangsu Province Hospital

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Variable analysis

independent variables

P53-shRNA-1
P53 shRNA-2
AMPKα1 shRNA

dependent variables

Expression of targeted protein (AMPKα1 and p53)

control variables

Scramble control shRNA lentiviral particles

controls

Positive control: Scramble control shRNA lentiviral particles
Negative control: Not explicitly mentioned

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