Immunoblot Analysis of T Cell Proteins

Whole cell lysates were collected from T cell samples as described previously (75 (link)). Antibodies for immunoblot analysis were Irp2 (IREB2; Thermo Fisher Scientific, PA5-19158), HFE (Invitrogen, PA5-37364), and β-actin (Cell Signaling Technology, 3700S). Blots were incubated with IRDye anti-rabbit or anti-mouse secondary antibodies (LI-COR) and imaged using the Odyssey CLx instrument. Band intensity quantification was conducted with Studio Lite software (LI-COR), and total protein signals were normalized to the β-actin signal for each sample.

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Voss K., Sewell A.E., Krystofiak E.S., Gibson-Corley K.N., Young A.C., Basham J.H., Sugiura A., Arner E.N., Beavers W.N., Kunkle D.E., Dickson M.E., Needle G.A., Skaar E.P., Rathmell W.K., Ormseth M.J., Major A.S, & Rathmell J.C. (2023). Elevated transferrin receptor impairs T cell metabolism and function in systemic lupus erythematosus. Science immunology, 8(79), eabq0178.

Publication 2023

β-actin IRDye anti-rabbit or anti-mouse secondary antibodies Studio Lite software

Corresponding Organization : Vanderbilt University Medical Center

Other organizations : Vanderbilt University, United States Department of Veterans Affairs, VA Tennessee Valley Healthcare System

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Variable analysis

independent variables

T cell samples

dependent variables

Irp2 (IREB2) protein levels
HFE protein levels
β-actin protein levels

control variables

Whole cell lysates collected from T cell samples as described previously (75)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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