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tra12015-sup-0001-FigureS1.docWord document1403K Figure S1: Overexpression of ORP5 in VPS4 knockdown cells. (Associated with Figure 2; signifies that ORP5 overexpression had little effect on cholesterol accumulation caused by VPS4 depletion) HeLa cells were treated with siVps4A and siVps4B for 24 h and then transfected with cDNAs encoding GFP-ORP5 or GFP-ORP5LBD for 24 h, followed by filipin staining for free cholesterol. Representative fluorescent images are shown. Arrow heads indicate ORP5 or ORP5LBD expressing cells. Bar = 10 µm.
tra12015-sup-0002-FigureS2.docWord document822K Figure S2: Overexpression of STARD4 in VPS4 knockdown cells. (Associated with Figure 2; signifies that STARD4 overexpression had little effect on cholesterol accumulation caused by VPS4 depletion) HeLa cells were treated with indicated siRNAs for 24 h and then transfected with cDNAs encoding Flag-STARD4 for 24 h, followed by immunofluorescence and filipin staining. Representative fluorescent images are shown. Arrow heads indicate Flag-STARD4 expressing cells. Bar = 10 µm.
tra12015-sup-0003-FigureS3.docWord document370K Figure S3: Uptake of BODIPY-LDL. (Associated with Figure 3; signifies that LDL uptake was not affected by VPS4 depletion) siRNA treated HeLa cells were incubated with BODIPY-LDL followed by measuring BODIPY signal presented in total cell lysate. Data were normalized to total proteins. BODIPY-LDL uptake in VPS4 knockdown cells was presented relative to that in control cells.
tra12015-sup-0004-FigureS4.docWord document2772K Figure S4: The effects of depleting VPS4 and ESCRT-III subunits on nuclear morphology and on endosomal cholesterol. (Associated with Figures 7 and 8; signifies that VPS4 has a function distinct from that of the ESCRT-III complex by its essential role in endosomal cholesterol transport) HeLa cells were transfected with indicated siRNAs for 48 h and then processing for Hoechst 33342 (Invitrogen) staining for nucleus or filipin staining for free cholesterol. Representative fluorescent images are shown. Arrow heads indicate abnormal nuclei. Bar = 10 µm.

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