Incubation:Article Title: ABCE1-dependent translational control links Fe-S cluster biogenesis to parasite growth and lipid homeostasis in Toxoplasma gondii
Article Snippet: .. HFF cells were pretreated with oleic acid (HY-N1446, MedChemExpress) at 400 μM and BODIPY 493/503 at 10 μM for 18h, washed five times with PBS and then incubated in complete DMEM medium for 30 min. Then, the cells were incubated with oleic acid at 200 μM and infected with cKD HA-ABCE1 tachyzoites either in the presence or absence of ATc for 48 h or with microtubule-disrupting agent oryzalin (HY-147092, MedChemExpress) at 2.5 μM for 48 h. Upon treatment with 2.5 μM of oryzalin for 48 h, the compound was removed by washes in HBSS and the parasites were then maintained in complete medium for 48 h. The parasites were fixed and permeabilized according to the IFA protocol, after which the LDs were stained with BODIPY493/503 for 30 min. ..
Infection:Article Title: ABCE1-dependent translational control links Fe-S cluster biogenesis to parasite growth and lipid homeostasis in Toxoplasma gondii
Article Snippet: .. HFF cells were pretreated with oleic acid (HY-N1446, MedChemExpress) at 400 μM and BODIPY 493/503 at 10 μM for 18h, washed five times with PBS and then incubated in complete DMEM medium for 30 min. Then, the cells were incubated with oleic acid at 200 μM and infected with cKD HA-ABCE1 tachyzoites either in the presence or absence of ATc for 48 h or with microtubule-disrupting agent oryzalin (HY-147092, MedChemExpress) at 2.5 μM for 48 h. Upon treatment with 2.5 μM of oryzalin for 48 h, the compound was removed by washes in HBSS and the parasites were then maintained in complete medium for 48 h. The parasites were fixed and permeabilized according to the IFA protocol, after which the LDs were stained with BODIPY493/503 for 30 min. ..
Immunofluorescence:Article Title: ABCE1-dependent translational control links Fe-S cluster biogenesis to parasite growth and lipid homeostasis in Toxoplasma gondii
Article Snippet: .. HFF cells were pretreated with oleic acid (HY-N1446, MedChemExpress) at 400 μM and BODIPY 493/503 at 10 μM for 18h, washed five times with PBS and then incubated in complete DMEM medium for 30 min. Then, the cells were incubated with oleic acid at 200 μM and infected with cKD HA-ABCE1 tachyzoites either in the presence or absence of ATc for 48 h or with microtubule-disrupting agent oryzalin (HY-147092, MedChemExpress) at 2.5 μM for 48 h. Upon treatment with 2.5 μM of oryzalin for 48 h, the compound was removed by washes in HBSS and the parasites were then maintained in complete medium for 48 h. The parasites were fixed and permeabilized according to the IFA protocol, after which the LDs were stained with BODIPY493/503 for 30 min. ..
Staining:Article Title: ABCE1-dependent translational control links Fe-S cluster biogenesis to parasite growth and lipid homeostasis in Toxoplasma gondii
Article Snippet: .. HFF cells were pretreated with oleic acid (HY-N1446, MedChemExpress) at 400 μM and BODIPY 493/503 at 10 μM for 18h, washed five times with PBS and then incubated in complete DMEM medium for 30 min. Then, the cells were incubated with oleic acid at 200 μM and infected with cKD HA-ABCE1 tachyzoites either in the presence or absence of ATc for 48 h or with microtubule-disrupting agent oryzalin (HY-147092, MedChemExpress) at 2.5 μM for 48 h. Upon treatment with 2.5 μM of oryzalin for 48 h, the compound was removed by washes in HBSS and the parasites were then maintained in complete medium for 48 h. The parasites were fixed and permeabilized according to the IFA protocol, after which the LDs were stained with BODIPY493/503 for 30 min. ..
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