All the DNA constructs and antibodies used in this study are listed in Supplementary Tables I and II , respectively. Most of the chemicals used in this study were purchased from Sigma-Aldrich. Alexa-fluor-conjugated-dextran, LysoTracker, LysoSensor dyes, EGF, and DAPI were purchased from Molecular Probes. The SiR-Lysosome Kit was purchased from Cytoskeleton, Inc., and the self-quenched BODIPY-FL conjugate of BSA was purchased from BioVision. Water-based ferrofluid (EMG 508) was purchased from Ferrotec Corporation. The chemical compounds, ML-098 and CID-1067700, were purchased from Cayman Chemicals.
The siRNA oligos for gene silencing were purchased from Dharmacon (Horizon Discovery) and prepared according to the manufacturer’s instructions. Transient transfection of siRNAs was performed using the DharmaFECT 1 reagent according to the manufacturer’s instructions. The following siRNAs were used in this study: control siRNA, 5’-TGGTTTACATGTCGACTAA-3’; ORF3a siRNA, 5’-GAGAATCTTCACAATTGGAACTGTA-3’56 (link); Rab7 siRNA, 5’-CTAGATAGCTGGAGAGATG-3’ and Vps39 siRNA 5’-CATTGCAGTGTTGCCTCGATATG-3’.
The siRNA oligos for gene silencing were purchased from Dharmacon (Horizon Discovery) and prepared according to the manufacturer’s instructions. Transient transfection of siRNAs was performed using the DharmaFECT 1 reagent according to the manufacturer’s instructions. The following siRNAs were used in this study: control siRNA, 5’-TGGTTTACATGTCGACTAA-3’; ORF3a siRNA, 5’-GAGAATCTTCACAATTGGAACTGTA-3’56 (link); Rab7 siRNA, 5’-CTAGATAGCTGGAGAGATG-3’ and Vps39 siRNA 5’-CATTGCAGTGTTGCCTCGATATG-3’.
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