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| Status |
Public on Nov 06, 2019 |
| Title |
A lysosome-plasma membrane-sphingolipid axis linking lysosomal storage to cell growth arrest |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
We used human fibroblasts loaded with sucrose as a simple model of lysosomal accumulation, and we verified by high-throughput RNA sequencing which processes were altered by the treatment at two different time points. Moreover, we hypothesized that ceramide production due to the ectopic catabolism of plasma-membrane glycosphingolipids is involved in cell cycle arrest upon sucrose treatment. To test this hypothesis, we treated sucrose-loaded fibroblasts with two specific inhibitors for the β-glucosidases GBA1 and GBA2, and analyzed the cell responses using high-throughput RNA sequence.
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| Overall design |
Poly(A) RNA capture followed by multiparallel sequencing performed in human skin fibroblasts cultured in presence or absence of sucrose. Three series of experiments have been performed. Series nr 1 includes samples cultured in presence or absence of sucrose for 14 days, and was performed on 3 different fibroblast cell lines (F1, L37 and L40, biological replicates), each in duplicate or triplicate, N=14. Series nr 2 includes samples cultured in presence or absence of sucrose for 2 days, and was performed on one fibroblast cell lines (L40) in triplicate, N=6. Series nr 3 includes samples cultured in presence of sucrose for 14 days, and treated for the last 48 hours with CBE and AMP-DNM, which specifically inhibits β-glucosidases GBA1 and GBA2 (N=3).
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| Contributor(s) |
Soldà G, Straniero L, Samarani M, Aureli M |
| Citation(s) |
29746165 |
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| Submission date |
Nov 15, 2017 |
| Last update date |
Feb 06, 2020 |
| Contact name |
Giulia Solda' |
| E-mail(s) |
giulia.solda@hunimed.eu
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| Organization name |
Humanitas University
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| Street address |
via Rita Levi Montalcini 4
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| City |
Pieve Emanuele-Milan |
| ZIP/Postal code |
20072 |
| Country |
Italy |
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| Platforms (1) |
| GPL18573 |
Illumina NextSeq 500 (Homo sapiens) |
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| Samples (23)
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| Relations |
| BioProject |
PRJNA418450 |
| SRA |
SRP125030 |
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