Immunoblot Analysis of Cellular Signaling

The immunoblot analysis was performed as described previously [44 (link)], using SuperSignal West Pico Substrate (Pierce, Rockford, IL, USA) and Western BLoT Hyper HRP Substrate (Takara Bio Inc., Shiga, Japan) according to the manufacturers’ instructions. Images were analysed using UN-SCAN-Itgel Automated Digitizing System software (Version 5.1 for Windows, Silk Scientific Inc., Orem, UT, USA). The antibodies to the following chemicals were used: eIF4E, p-eIF4E (Ser209), MNK1, MNK2, vimentin, N-cadherin, and E-cadherin. β-actin was used as a loading control.

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Ichiyanagi O., Ito H., Naito S., Kabasawa T., Kanno H., Narisawa T., Ushijima M., Kurota Y., Ozawa M., Sakurai T., Nishida H., Kato T., Yamakawa M, & Tsuchiya N. (2019). Impact of eIF4E phosphorylation at Ser209 via MNK2a on tumour recurrence after curative surgery in localized clear cell renal cell carcinoma. Oncotarget, 10(40), 4053-4068.

Publication 2019

Western BLoT Hyper HRP Substrate UN-SCAN-Itgel Automated Digitizing System software

Actin Antibodies E cadherin Eif4e Immunoblot analysis Mnk1 Mnk2 N cadherin Scan Silk Vimentin Western blot

Corresponding Organization : Yamagata University

Other organizations : Kihoku hospital

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Variable analysis

independent variables

Antibodies to the following chemicals: eIF4E, p-eIF4E (Ser209), MNK1, MNK2, vimentin, N-cadherin, and E-cadherin

dependent variables

Immunoblot analysis

control variables

β-actin as a loading control

controls

Positive control: None specified
Negative control: None specified

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