DOI: 10.5061/dryad.p253c6f
title: Data from: Apolipoprotein L-1 renal risk variants form active channels at the plasma membrane driving cytotoxicity

description: Recently evolved alleles of Apolipoprotein L-1 (APOL1) provide increased protection against African trypanosome parasites while also significantly increasing the risk of developing kidney disease in humans. APOL1 protects against trypanosome infections by forming ion channels within the parasite, causing lysis. While the correlation to kidney disease is robust, there is little consensus concerning the underlying disease mechanism. We show in human cells that the APOL1 renal risk variants have a population of active channels at the plasma membrane, which results in an influx of both Na+ and Ca2+. We propose a model wherein APOL1 channel activity is the upstream event causing cell death, and that the activate-state, plasma membrane-localized channel represents the ideal drug target to combat APOL1-mediated kidney disease.
description: Figure_1C-FT293_timecouresesCell death was measured in FlpIn TREX 293 cells expressing three major variants of APOL1, G0, G1, and G2. Enclosed is the analyzed percent cytotoxicity measured via lactate dehydrogenase assay as well as the statistical test for each biological replicate.Figure_2C-FT293_CaCl2_celldeathMeasuring the effects of increased Ca2+ in the cell culture media on the cytotoxicity of the kidney disease risk variants of APOL1. Enclosed is the analyzed percent cytotoxicity measured via lactate dehydrogenase assay as well as the statistical test for each biological replicate.Figure_2D-Fluorescence-traces-of-representative-cellsFluorescence traces from GCaMP6f transfected FlpIn TREX 293 cells expressing the APOL1 variants. Enclosed is the analyzed GCaMP6f ∆F/F0 of the representative cells depicted in Fig 2D.Figure_2E-analyzed-cells-representative-timepointsThis files contains the calculated GCaMP6f ∆F/F0 of each cell analyzed in Fig. 2E.Figure_2E-raw-tracking-dataThis file contains the raw tracking data (using TrackMate) of each field of view imaged and analyzed. Cells imaged were GCaMP6f transfected, APOL1 variants expressing FlpIn TREX 293 cells. Each tab within the excel file contains the raw tracking data for a single condition (Genotype +/- APOL1 expression). A summary tab is also included.Figure_3B-RUSH_APOL1_cytotoxicityIn order to determine if APOL1 cytotoxicity required trafficking from the ER, HEK293 cells were transfected with RUSH-APOL1 vectors, and the cytotoxicity of cells treated with (releasing APOL1 from the ER) or without biotin were compared. Enclosed is the analyzed percent cytotoxicity measured via lactate dehydrogenase assay as well as the statistical test for each biological replicate.Figure_3C-Fluorescence-traces-of-representative-cellsThe GCaMP6f ∆F/F0 of each representative cell depicted in Figure 3C.Figure_3D-raw-tracking-dataRaw tracking data (using TrackMate) of each cell analyzed in Figure 3D.Figure_3D-analyzed-cells-representative-timepointsThe calculated GCaMP6f ∆F/F0 of each cell analyzed from Video 2 and graphed in Figure 3D.Figure 4D and E - Statistical analysisStatistical analysis of the APOL1 cell surface quantification.Figure_4C-Raw_APOL1_Intensity_at_SurfaceThe raw signal intensity of APOL1 at the plasma membrane, calculated from IF images. The signal intensity of each analyzed cell from 3 replicates is depicted in this file. Further calculations were performed in Figures 4D and E in order to better visualize the data.Figure_5B-Ammonium_ChlorideCytotoxicity of RUSH-APOL1 transfected cells treated with or without ammonium chloride. Enclosed is the analyzed percent cytotoxicity measured via lactate dehydrogenase assay as well as the statistical test for each biological replicate.Figure_5A-Acid_NeutralCytotoxicity of RUSH-APOL1 transfected cells with or without acid treatment. Enclosed is the analyzed percent cytotoxicity measured via lactate dehydrogenase assay as well as the statistical test for each biological replicate.