Inducing APOL1 expression in kidney organoids

Day 24 G0 and G1 kidney organoids in identically plated wells of a 24-well plate were treated with IFN-γ (25 ng/ml; PeproTech) for 24 hours to induce APOL1 expression. This dose approximates previous in vitro IFN-γ doses used for macrophage activation (22 (link),23 (link)). Endoplasmic reticulum (ER) stress was induced by adding 5 μM tunicamycin (Tocris) for 24 hours.

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Liu E., Radmanesh B., Chung B.H., Donnan M.D., Yi D., Dadi A., Smith K.D., Himmelfarb J., Li M., Freedman B.S, & Lin J. (2020). Profiling APOL1 Nephropathy Risk Variants in Genome-Edited Kidney Organoids with Single-Cell Transcriptomics. Kidney360, 1(3), 203-215.

Publication 2020

IFN-γ tunicamycin

Apol1 Endoplasmic reticulum stress Ifn γ Kidney Macrophage activation Organoids Tunicamycin

Corresponding Organization : Jesse Brown VA Medical Center

Other organizations : University of Washington

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Variable analysis

independent variables

Treatment with IFN-γ (25 ng/ml) for 24 hours
Induction of endoplasmic reticulum (ER) stress by adding 5 μM tunicamycin for 24 hours

dependent variables

APOL1 expression

control variables

Day 24 G0 and G1 kidney organoids in identically plated wells of a 24-well plate

controls

Positive control: Previous in vitro IFN-γ doses used for macrophage activation (22, 23)
Negative control: Not explicitly mentioned

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