Cell culture
By Emma Sherrell onThis protocol describes routine cell culture maintenance as described in Adriaenssens & Nguyen et al., 2023 (BioRxiv).
Plasmid construction and Gibson cloning
By Emma Sherrell onThis protocol describes plasmid construction as described in Adriaenssens & Nguyen et al., 2023 (BioRxiv).
Immunofluorescence assay (IFA)
By Emma Sherrell onProtocol for performing an immunofluorescence assay with fixed HeLa cells.
qRT-PCR sample preparation
By Emma Sherrell onProtocol for qRT-PCR sample preparation for analysis using a RotorGeneQ machine (Qiagen).
Analyzing oxygen consumption of isolated mitochondria using the Seahorse XFe96 analyzer
By Emma Sherrell onProtocol for analyzing oxygen consumption of isolated mitochondria using the Seahorse XFe96 analyzer.
Analyzing cellular ATP levels
By Emma Sherrell onProtocol for analysis of cellular ATP levels using the Promega Mitochondrial ToxGlo Kit.
Measuring mitophagy via FACS with mtKeima reporter
By Emma Sherrell onPreparation of samples for measuring mitophagy levels using mtKeima reporter by fluorescence activated cell sorting (FACS).
Calculating mitochondrial protein solubility
By Emma Sherrell onProtocol for generating solubility calculations for mitochondrial proteins from starting 'soluble' and 'insoluble' fraction mass spectrometry data, using MaxQuant and Perseus software pipelines.
Reverse-phase high pH fractionation (using Thermo Fisher, Cat# 84868)
By Emma Sherrell onThis protocol uses the Pierce™ High pH Reversed-Phase Peptide Fractionation Kit (Thermo Fisher, Cat# 84868).
Preparation of soluble and insoluble mitochondrial protein fractions for immunoblotting
By Emma Sherrell onPreparation of soluble and insoluble mitochondrial protein fractions from HeLa cells for immunoblotting.
Mitochondrial isolation from HeLa cells
By Emma Sherrell onMitochondrial isolation and quantification from HeLa cells.
Western blotting using the BioRad Criterion Blotter system
By Emma Sherrell onProtocol for performing an SDS-PAGE Western blot analysis using the BioRad Criterion Blotter system.
Preparing whole cell samples for immunoblot analysis
By Emma Sherrell onProtocol for preparation of HeLa cell lysates for immunoblot analysis.
Induction of starvation stress
By Emma Sherrell onInduction of starvation stress using EBSS in HeLa cells.
Mitophagy induction using Difereprone
By Emma Sherrell onMitophagy induction in HeLa cells using Difereprone (DFP).
Mitophagy induction using Oligomycin/Antimycin A
By Emma Sherrell onMitophagy induction in HeLa cells using Oligomycin/Antimycin A.
Generation of stable cell lines using viral infection
By Emma Sherrell onProtocol for generation and precipitation of retrovirus, and infection of HeLa cells to generate stable cell lines.
Salivary collection participant instructions using OMNIgene ORAL OM 501 device
By Emma Sherrell onSalivary collection participant instructions using OMNIgene ORAL OM 501 device.