Knockdown of ATP6V0A2 by shRNA

Oligonucleotides containing the siRNA-expressing sequence targeting ATP6V0A2 were annealed (shATPV0A2 top: 5′-GATCCCCGCAGCTTTGACGTGACCAACATTCGAAGAGTGTTGGTCACGTCAAAGCTGCTTTTTA-3′, shATP6V0A2 bottom: 5′-AGCTTAAAAAGCAGCTTTGACGTGACCAACACTCTTCGAATGTTGGTCACGTCAAAGCTGCGGG-3′), and cloned into the pSUPER.retro vector (OligoEngine, Seattle, WA). Preparation of viral supernatant and infection of target cells was performed as described26 (link). Transduced cells were selected with 2 μg/mL puromycin for 3 days.

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Udono M., Fujii K., Harada G., Tsuzuki Y., Kadooka K., Zhang P., Fujii H., Amano M., Nishimura S.I., Tashiro K., Kuhara S, & Katakura Y. (2015). Impaired ATP6V0A2 expression contributes to Golgi dispersion and glycosylation changes in senescent cells. Scientific Reports, 5, 17342.

Publication 2015

pSUPER.retro vector

Cells Infection Oligonucleotides Puromycin Sirna Vector

Corresponding Organization :

Other organizations : Kyushu University, Hokkaido University

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Variable analysis

independent variables

Oligonucleotides containing the siRNA-expressing sequence targeting ATP6V0A2

dependent variables

Transduced cells

control variables

Preparation of viral supernatant and infection of target cells (as described in the referenced publication)

controls

Positive control: Not specified
Negative control: Not specified

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