Modulating Autophagy and NF-κB in Invasion Assays

To inhibit lysosomal protease, 10 μg/mL E-64d (permeable form, Sigma, USA) dissolved in DMSO and 2.5 μg/mL pepstatin A (Sigma) in 10% acetic acid in methanol were used for 4 h. Rapamycin 1 μM (Sigma) was used to induce autophagy in invasion assays. Bafilomycin A1 0.1 μM (Sigma) was used to inhibit autophagy in invasion assays. To inhibit NF-κB activity, an NF-κB inhibitor (100 μg/mL, sc-3060, Santa Cruz, USA), which has been previously used to specifically inhibit NF-kB activity^{63 (link),64 (link)}, or the NF-κB control (sc-3061, Santa Cruz) was used.

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Oh S.Y., Hwang J.R., Choi M., Kim Y.M., Kim J.S., Suh Y.L., Choi S.J, & Roh C.R. (2020). Autophagy regulates trophoblast invasion by targeting NF-κB activity. Scientific Reports, 10, 14033.

Publication 2020

pepstatin A Rapamycin Bafilomycin A1 sc-3060 sc-3061

Acetic acid Assays Autophagy Bafilomycin a1 Dmso Inhibit Lysosomal Methanol Nf κb Nf κb inhibitor Pepstatin Permeable Protease Rapamycin Sc 3060

Corresponding Organization :

Other organizations : Samsung Medical Center, Sungkyunkwan University, Chung-Ang University Hospital

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Variable analysis

independent variables

E-64d (10 μg/mL)
Pepstatin A (2.5 μg/mL)
Rapamycin (1 μM)
Bafilomycin A1 (0.1 μM)
NF-κB inhibitor (100 μg/mL)
NF-κB control (sc-3061)

dependent variables

Invasion assays

control variables

DMSO used to dissolve E-64d
10% acetic acid in methanol used to dissolve Pepstatin A

positive control

NF-κB inhibitor (100 μg/mL)

negative control

NF-κB control (sc-3061)

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