Protein Expression Analysis via Western Blot

Cells were lysed in RIPA-SDS buffer, after which the proteins were resolved by
SDS-PAGE, transferred onto PVDF membranes, and probed with the relevant
antibodies as previously described9 (link). The following antibodies
were used for the experiments: anti-actin (Sigma); anti-SESN2 (Proteintech),
anti-NRF2, anti-CHOP and anti-p53 (Santa Cruz); and anti-phospho(T389)-p70S6K,
anti-phospho(S235/236)-S6, anti-phospho(S65)-4E-BP1, anti-phospho
IF2α(S51), anti-phospho(T172)-AMPKα,
anti-phospho(S79)-acetyl-CoA carboxylase, anti-phospho(S9)-GSK3β,
anti-phospho(T1462), anti-4E-BP1, anti-S6, anti-TSC2, anti-AMPKα,
anti-ATF4, anti-Bip, anti-p62, anti-LC3, anti-PARP, anti-PARP(cleaved),
anti-caspase3, and anti-caspase3(cleaved) (Cell Signaling Inc). Sesn1 antibodies
were previously described11 (link).

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Ding B., Parmigiani A., Divakaruni A.S., Archer K., Murphy A.N, & Budanov A.V. (2016). Sestrin2 is induced by glucose starvation via the unfolded protein response and protects cells from non-canonical necroptotic cell death. Scientific Reports, 6, 22538.

Publication 2016

anti-actin anti-SESN2 anti-NRF2 anti-CHOP anti-p53 anti-phospho(S65)-4E-BP1 anti-phospho(T172)-AMPKα anti-4E-BP1 anti-S6 anti-TSC2 anti-AMPKα anti-ATF4 anti-Bip anti-p62 anti-LC3 anti-PARP anti-caspase3

4e bp1 Acetyl coa carboxylase Actin Atf4 Buffer Caspase3 Cells Chop Gsk3β Membranes Nrf2 P70s6k Proteins Pvdf Ripa Tsc2

Corresponding Organization :

Other organizations : Virginia Commonwealth University, University of California, San Diego

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

Protein expression levels of actin, SESN2, NRF2, CHOP, p53, phospho(T389)-p70S6K, phospho(S235/236)-S6, phospho(S65)-4E-BP1, phospho(S51)-IF2α, phospho(T172)-AMPKα, phospho(S79)-acetyl-CoA carboxylase, phospho(S9)-GSK3β, phospho(T1462)-TSC2, 4E-BP1, S6, TSC2, AMPKα, ATF4, Bip, p62, LC3, PARP, cleaved PARP, caspase3, and cleaved caspase3

control variables

Cells were lysed in RIPA-SDS buffer
Proteins were resolved by SDS-PAGE and transferred onto PVDF membranes
Antibodies used for probing were as previously described in reference 9 (link) and reference 11 (link)

positive controls

Not specified

negative controls

Not specified

Annotations

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