S-BIAD144alpha

The glucosylceramide synthase inhibitor PDMP causes lyso-somal lipid accumulation and mTOR inactivation

Released 2021-05-31
Pia Hartwig, Doris Höglinger

Content

  • 223 images
We investigated subcellular sphingolipid distribution using a functionalized sphingosine analogue (pacSph) by confocal microscopy in control, PDMP and NB-DNJ (Miglustat) treated WT and SGPL1 knock-out cells (experimentA; channel 1: pacSph, channel 2: Lamp1). We studied lysosomal sphingolipid (pacSph) export using pulse-chase experiments in control and PDMP treated SGPL1 knock-out cells (experimentB; channel 1: pacSph, channel 2: Lamp1). We analyzed other lipids such as LBPA (immunofluorescence staining) and cholesterol (filipin staining) at the lysosome in control, PDMP and NB-DNJ (Miglustat) treated WT cells by microscopy (experimentC). We investigated a timeline of lipid accumulating effects arising from PDMP treatment by confocal microscopy. Therefore, we investigated LBPA (immunofluorescence staining in WT cells; channel 1: LBPA, channel 2: Lamp1), sphingolipids (using pacSph in SGPL1 knock-out cells; channel 1: pacSph, channel 2: Lamp1) and cholesterol (filipin staining in WT cells; channel 1: Filipin, channel 2: Lamp1) (experimentD). We analysed the timeline of mTOR (channel 1: mTOR, channel 2: Lamp1) and TFEB (channel 1: DAPI, channel 2: TFEB) subcellular translocation arising from PDMP treatment in WT cells by immunofluorescence microscopy (experimentE).

Study information

Organism
Homo sapiens (human)
Imaging type
confocal fluorescence microscopy
License
CC0

Viewable images

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IM1 experiments/experimentA_01_WT_PDMP.czi (1, 2, 1, 512, 512) Unavailable
IM2 experiments/experimentA_02_WT_PDMP.czi (1, 2, 1, 512, 512) Unavailable
IM3 experiments/experimentA_03_WT_PDMP.czi (1, 2, 1, 512, 512) Unavailable
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IM6 experiments/experimentA_06_WT_mock.czi (1, 2, 1, 512, 512) Unavailable
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IM10 experiments/experimentA_10_WT_Miglustat.czi (1, 2, 1, 512, 512) Unavailable
IM11 experiments/experimentA_11_WT_Miglustat.czi (1, 2, 1, 512, 512) Unavailable
IM12 experiments/experimentA_12_SGPL1_PDMP.czi (1, 2, 1, 512, 512) Unavailable
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IM20 experiments/experimentA_20_SGPL1_Miglustat.czi (1, 2, 1, 512, 512) Unavailable
IM23 experiments/experimentB_02_PDMP_0min.czi (1, 2, 1, 512, 512) Unavailable
IM50 experiments/experimentB_29_5min.czi (1, 2, 1, 512, 512) Unavailable
IM198 experiments/experimentE_mTOR_23_6h.czi (1, 2, 1, 512, 512) Unavailable
IM200 experiments/experimentE_mTOR_25_6h.czi (1, 2, 1, 512, 512) Unavailable

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