Western Blot Analysis of Antioxidant and EMT Markers

Cells were lysed with lysis buffer (P89901; Thermo Scientific) containing protease inhibitors (P8340; Sigma-Aldrich). Western blot analysis was performed as previously described 10 (link). The following primary antibodies were used: anti-Trx-1 (1: 1000), anti-G6PD (1:2000), and GAPDH (1:5000) from Abcam (Cambridge, UK); anti-E-cadherin (1:1000; Cell Signaling Technology, Beverly, MA), and anti-vimentin (1:5000; Santa Cruz, CA, USA). Protein band intensity was quantified using Image Lab Software (Bio-Rad Laboratories Inc., Berkeley, CA, USA).

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Lu F., Fang D., Li S., Zhong Z., Jiang X., Qi Q., Liu Y., Zhang W., Xu X., Liu Y., Zhu W, & Jiang L. (2022). Thioredoxin 1 supports colorectal cancer cell survival and promotes migration and invasion under glucose deprivation through interaction with G6PD. International Journal of Biological Sciences, 18(14), 5539-5553.

Publication 2022

P89901 P8340 anti-Trx-1 anti-G6PD GAPDH anti-E-cadherin anti-vimentin Image Lab Software

Antibodies Buffer Cadherin 1 Cells G6pd Gapdh Protease inhibitors Protein Trx 1 Vimentin Western blot analysis

Corresponding Organization : First Affiliated Hospital of Wenzhou Medical University

Other organizations : Nanjing Medical University

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Variable analysis

independent variables

Lysis buffer (P89901; Thermo Scientific) containing protease inhibitors (P8340; Sigma-Aldrich)

dependent variables

Protein band intensity

control variables

Western blot analysis protocol as previously described 10 (link)
Primary antibodies: anti-Trx-1 (1:1000), anti-G6PD (1:2000), GAPDH (1:5000) from Abcam (Cambridge, UK); anti-E-cadherin (1:1000; Cell Signaling Technology, Beverly, MA), and anti-vimentin (1:5000; Santa Cruz, CA, USA)
Protein band intensity quantification using Image Lab Software (Bio-Rad Laboratories Inc., Berkeley, CA, USA)

controls

Positive controls: Not specified
Negative controls: Not specified

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