FoxO suppresses endoplasmic reticulum stress to inhibit growth of Tsc1-deficient tissues under nutrient restriction
- Institute of Molecular Systems Biology, ETH Zürich, Switzerland
Figures
Figure 1 with 2 supplements
Transcriptomics analysis to identify FoxO targets in Tsc1 mutant cells.
(A) Schematic of the experimental setup to generate temporal knockdown of foxo in wild-type or Tsc1 mutant clones, and isolation of single clones using LCM at 108 hr after egg laying (AEL) from …
Figure 1—figure supplement 1
Validation of RNA-seq data.
(A) Multidimensional scaling plot of log counts per million (CPM) of ctrl and Tsc1 mutant cells, with or without foxo knockdown, from different food conditions. (B) foxo expression levels in …
Figure 1—figure supplement 2
Contamination status of isolated clones.
(A) Representative images of isolated Tsc1Q87X and Tsc1Q87X foxoRi clones. Blue dashed lines mark the GFP-negative clones from which the selected clones (outlined in yellow) were isolated. Scale bar …
Figure 2 with 2 supplements
In vivo RNAi assay to validate downregulated genes in Tsc1 mutant cells upon foxo knockdown.
(A) log fold change (FC) of downregulated genes in Tsc1 mutant cells with foxo knockdown versus Tsc1 mutant cells under NR. (B) Percentage change in adult eye size upon knockdown of candidate genes …
Figure 2—figure supplement 1
In vivo RNAi assay of downregulated candidates with strong Tsc1 knockdown and representative phenotypes of top hits.
(A) Percentage eclosion of flies with co-knockdown of candidate with Tsc1 under NR. Numbers above bars indicate the number of eclosed flies for each cross. (B) Representative images of eyes from …
Figure 2—figure supplement 2
Loss of CG6766 enhances ER stress in Tsc1 mutant cells.
(A) Calnexin 99A staining of eye imaginal discs with control (iso) or CG6766 mutant clones dissected from larvae raised under normal feeding conditions. Clones are marked by the absence of GFP, and …
Figure 3
Upregulation of ER stress increases Tsc1 knockdown overgrowth.
(A) Schematic depicting involvement of genes tested in (B), (C) and (D) in different ER stress regulation mechanisms according to the KEGG database. Blue, green and orange denote components of ERAD, …
Figure 4 with 1 supplement
Loss of foxo enhances ER stress in Tsc1 mutant cells.
(A) Calnexin 99A staining of eye imaginal discs with ctrl, foxo, Tsc1 or Tsc1 foxo mutant clones dissected from larvae raised under NR. Clones are marked by the absence of GFP; DAPI stains nuclei. …
Figure 4—figure supplement 1
FoxO suppresses Ire1 pathway activation in Tsc1 mutant cells under NR.
(A–B) GFP staining to detect Xbp1-EGFP of eye imaginal discs with ctrl, foxo, Tsc1 or Tsc1 foxo knockdown clones dissected from larvae raised under (A) normal food or (B) NR. Clones are marked by …
Author response image 1
GFP expression levels in Tsc1 mutant clones with and without foxo knockdown isolated from larvae raised under normal feeding conditions.
Data are represented as mean ± SD.
Author response image 2
Additional files
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Supplementary file 1
Key Resources Table.
- https://cdn.elifesciences.org/articles/53159/elife-53159-supp1-v2.docx
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Supplementary file 2
Primer sequences for dsRNA synthesis and qPCR.
- https://cdn.elifesciences.org/articles/53159/elife-53159-supp2-v2.docx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/53159/elife-53159-transrepform-v2.pdf
