Western Blot Analysis of UPR Pathway Proteins

Mouse spheres or tumor tissues lysates were used for Western blots as described [49 (link)]. For immunoblotting, we used the following antibodies: anti-mouse PERK (#3192 S), p-PERK (#3179 S), IRE1α (#43294 S), ATF6 (#65880), p-eIF2α (#3398 S), ATF4 (#11815 S), ERK1/2 (#4695 S), p-ERK (#5726 S), CHOP (#5554 S), PARP (9542 S), XPO-1 (#46249), lamin B1 (#12586 S), GAPDH (#3683 S), β-actin (#5125 S) were from Cell Signaling Technology (Danvers, MA), p21 (#sc-6246) was from Santa Cruz Biotechnology, Santa Cruz, CA),and p-p21 (#E-AB-20947) was from Elabscience (Huston, TX). Antibody binding to the membrane was visualized using a chemiluminescent detection system (ECL, Millipore Sigma).

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Hassett D.J., Alsabbagh E., Parvatiyar K., Howell M.L., Wilmott R.W, & Ochsner U.A. (2000). A Protease-Resistant Catalase, KatA, Released upon Cell Lysis during Stationary Phase Is Essential for Aerobic Survival of a Pseudomonas aeruginosa oxyR Mutant at Low Cell Densities. Journal of Bacteriology, 182(16), 4557-4563.

Publication 2023

anti-mouse PERK p-PERK IRE1α p-eIF2α ERK1/2 p-ERK lamin B1 GAPDH β-actin ECL

Actin Antibodies Antibody Atf4 Atf6 Chop Erk1 Gapdh Ire1α Lamin b1 Membrane Mouse Tissues Tumor Western blots

Corresponding Organization :

Other organizations : Cincinnati Children's Hospital Medical Center, University of Cincinnati, The University of Texas Health Science Center at San Antonio, The University of Texas Health Science Center at Houston, University of Cincinnati Medical Center

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Variable analysis

independent variables

Mouse spheres or tumor tissues lysates

dependent variables

Protein expression levels of PERK, p-PERK, IRE1α, ATF6, p-eIF2α, ATF4, ERK1/2, p-ERK, CHOP, PARP, XPO-1, lamin B1, GAPDH, β-actin, p21, p-p21

control variables

Not explicitly mentioned

controls

Positive control: Not specified
Negative control: Not specified

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