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Developmental depletion <t>of</t> <t>Tet</t> causes anterior-specific loss of ISCs and reduces adult lifespan. (A) Representative confocal images of whole midguts from control ( Luc RNAi ) and Tet RNAi adult flies driven by esg-Gal4, <t>UAS-nGFP</t> . Loss of ISCs/EBs (green) is restricted to the anterior midgut. (B) High-magnification images of the anterior and posterior midgut compartments across three independent Tet RNAi lines. (C) Quantification of ISC/EB percentage (GFP+/DAPI) in the anterior and posterior midgut. (D) Confirmation of ISC loss using the endogenous Tet-Gal4 T2A driver crossed with UAS-mCherry.nls and stained for the ISC marker Delta (green). (E) Quantification of ISC percentage (Delta+/DAPI) in the anterior midgut. (F) Survival curve of adult flies exposed to paraquat-induced oxidative stress over 48 hours. Tet RNAi n=54, Luciferase RNAi n=53. (G) Lifespan analysis comparing control and Tet RNAi adult flies. Tet RNAi #1 n=247, Tet RNAi #2 n=118, Luciferase RNAi n=240. Lifespans were analyzed with OASIS 2. Bars represent mean ± SD. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001. p values were from unpaired Student’s t-test.
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Developmental depletion <t>of</t> <t>Tet</t> causes anterior-specific loss of ISCs and reduces adult lifespan. (A) Representative confocal images of whole midguts from control ( Luc RNAi ) and Tet RNAi adult flies driven by esg-Gal4, <t>UAS-nGFP</t> . Loss of ISCs/EBs (green) is restricted to the anterior midgut. (B) High-magnification images of the anterior and posterior midgut compartments across three independent Tet RNAi lines. (C) Quantification of ISC/EB percentage (GFP+/DAPI) in the anterior and posterior midgut. (D) Confirmation of ISC loss using the endogenous Tet-Gal4 T2A driver crossed with UAS-mCherry.nls and stained for the ISC marker Delta (green). (E) Quantification of ISC percentage (Delta+/DAPI) in the anterior midgut. (F) Survival curve of adult flies exposed to paraquat-induced oxidative stress over 48 hours. Tet RNAi n=54, Luciferase RNAi n=53. (G) Lifespan analysis comparing control and Tet RNAi adult flies. Tet RNAi #1 n=247, Tet RNAi #2 n=118, Luciferase RNAi n=240. Lifespans were analyzed with OASIS 2. Bars represent mean ± SD. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001. p values were from unpaired Student’s t-test.
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Developmental depletion <t>of</t> <t>Tet</t> causes anterior-specific loss of ISCs and reduces adult lifespan. (A) Representative confocal images of whole midguts from control ( Luc RNAi ) and Tet RNAi adult flies driven by esg-Gal4, <t>UAS-nGFP</t> . Loss of ISCs/EBs (green) is restricted to the anterior midgut. (B) High-magnification images of the anterior and posterior midgut compartments across three independent Tet RNAi lines. (C) Quantification of ISC/EB percentage (GFP+/DAPI) in the anterior and posterior midgut. (D) Confirmation of ISC loss using the endogenous Tet-Gal4 T2A driver crossed with UAS-mCherry.nls and stained for the ISC marker Delta (green). (E) Quantification of ISC percentage (Delta+/DAPI) in the anterior midgut. (F) Survival curve of adult flies exposed to paraquat-induced oxidative stress over 48 hours. Tet RNAi n=54, Luciferase RNAi n=53. (G) Lifespan analysis comparing control and Tet RNAi adult flies. Tet RNAi #1 n=247, Tet RNAi #2 n=118, Luciferase RNAi n=240. Lifespans were analyzed with OASIS 2. Bars represent mean ± SD. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001. p values were from unpaired Student’s t-test.
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Developmental depletion <t>of</t> <t>Tet</t> causes anterior-specific loss of ISCs and reduces adult lifespan. (A) Representative confocal images of whole midguts from control ( Luc RNAi ) and Tet RNAi adult flies driven by esg-Gal4, <t>UAS-nGFP</t> . Loss of ISCs/EBs (green) is restricted to the anterior midgut. (B) High-magnification images of the anterior and posterior midgut compartments across three independent Tet RNAi lines. (C) Quantification of ISC/EB percentage (GFP+/DAPI) in the anterior and posterior midgut. (D) Confirmation of ISC loss using the endogenous Tet-Gal4 T2A driver crossed with UAS-mCherry.nls and stained for the ISC marker Delta (green). (E) Quantification of ISC percentage (Delta+/DAPI) in the anterior midgut. (F) Survival curve of adult flies exposed to paraquat-induced oxidative stress over 48 hours. Tet RNAi n=54, Luciferase RNAi n=53. (G) Lifespan analysis comparing control and Tet RNAi adult flies. Tet RNAi #1 n=247, Tet RNAi #2 n=118, Luciferase RNAi n=240. Lifespans were analyzed with OASIS 2. Bars represent mean ± SD. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001. p values were from unpaired Student’s t-test.
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Developmental depletion <t>of</t> <t>Tet</t> causes anterior-specific loss of ISCs and reduces adult lifespan. (A) Representative confocal images of whole midguts from control ( Luc RNAi ) and Tet RNAi adult flies driven by esg-Gal4, <t>UAS-nGFP</t> . Loss of ISCs/EBs (green) is restricted to the anterior midgut. (B) High-magnification images of the anterior and posterior midgut compartments across three independent Tet RNAi lines. (C) Quantification of ISC/EB percentage (GFP+/DAPI) in the anterior and posterior midgut. (D) Confirmation of ISC loss using the endogenous Tet-Gal4 T2A driver crossed with UAS-mCherry.nls and stained for the ISC marker Delta (green). (E) Quantification of ISC percentage (Delta+/DAPI) in the anterior midgut. (F) Survival curve of adult flies exposed to paraquat-induced oxidative stress over 48 hours. Tet RNAi n=54, Luciferase RNAi n=53. (G) Lifespan analysis comparing control and Tet RNAi adult flies. Tet RNAi #1 n=247, Tet RNAi #2 n=118, Luciferase RNAi n=240. Lifespans were analyzed with OASIS 2. Bars represent mean ± SD. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001. p values were from unpaired Student’s t-test.
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Developmental depletion <t>of</t> <t>Tet</t> causes anterior-specific loss of ISCs and reduces adult lifespan. (A) Representative confocal images of whole midguts from control ( Luc RNAi ) and Tet RNAi adult flies driven by esg-Gal4, <t>UAS-nGFP</t> . Loss of ISCs/EBs (green) is restricted to the anterior midgut. (B) High-magnification images of the anterior and posterior midgut compartments across three independent Tet RNAi lines. (C) Quantification of ISC/EB percentage (GFP+/DAPI) in the anterior and posterior midgut. (D) Confirmation of ISC loss using the endogenous Tet-Gal4 T2A driver crossed with UAS-mCherry.nls and stained for the ISC marker Delta (green). (E) Quantification of ISC percentage (Delta+/DAPI) in the anterior midgut. (F) Survival curve of adult flies exposed to paraquat-induced oxidative stress over 48 hours. Tet RNAi n=54, Luciferase RNAi n=53. (G) Lifespan analysis comparing control and Tet RNAi adult flies. Tet RNAi #1 n=247, Tet RNAi #2 n=118, Luciferase RNAi n=240. Lifespans were analyzed with OASIS 2. Bars represent mean ± SD. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001. p values were from unpaired Student’s t-test.
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Developmental depletion <t>of</t> <t>Tet</t> causes anterior-specific loss of ISCs and reduces adult lifespan. (A) Representative confocal images of whole midguts from control ( Luc RNAi ) and Tet RNAi adult flies driven by esg-Gal4, <t>UAS-nGFP</t> . Loss of ISCs/EBs (green) is restricted to the anterior midgut. (B) High-magnification images of the anterior and posterior midgut compartments across three independent Tet RNAi lines. (C) Quantification of ISC/EB percentage (GFP+/DAPI) in the anterior and posterior midgut. (D) Confirmation of ISC loss using the endogenous Tet-Gal4 T2A driver crossed with UAS-mCherry.nls and stained for the ISC marker Delta (green). (E) Quantification of ISC percentage (Delta+/DAPI) in the anterior midgut. (F) Survival curve of adult flies exposed to paraquat-induced oxidative stress over 48 hours. Tet RNAi n=54, Luciferase RNAi n=53. (G) Lifespan analysis comparing control and Tet RNAi adult flies. Tet RNAi #1 n=247, Tet RNAi #2 n=118, Luciferase RNAi n=240. Lifespans were analyzed with OASIS 2. Bars represent mean ± SD. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001. p values were from unpaired Student’s t-test.
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Developmental depletion <t>of</t> <t>Tet</t> causes anterior-specific loss of ISCs and reduces adult lifespan. (A) Representative confocal images of whole midguts from control ( Luc RNAi ) and Tet RNAi adult flies driven by esg-Gal4, <t>UAS-nGFP</t> . Loss of ISCs/EBs (green) is restricted to the anterior midgut. (B) High-magnification images of the anterior and posterior midgut compartments across three independent Tet RNAi lines. (C) Quantification of ISC/EB percentage (GFP+/DAPI) in the anterior and posterior midgut. (D) Confirmation of ISC loss using the endogenous Tet-Gal4 T2A driver crossed with UAS-mCherry.nls and stained for the ISC marker Delta (green). (E) Quantification of ISC percentage (Delta+/DAPI) in the anterior midgut. (F) Survival curve of adult flies exposed to paraquat-induced oxidative stress over 48 hours. Tet RNAi n=54, Luciferase RNAi n=53. (G) Lifespan analysis comparing control and Tet RNAi adult flies. Tet RNAi #1 n=247, Tet RNAi #2 n=118, Luciferase RNAi n=240. Lifespans were analyzed with OASIS 2. Bars represent mean ± SD. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001. p values were from unpaired Student’s t-test.
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Image Search Results


Developmental depletion of Tet causes anterior-specific loss of ISCs and reduces adult lifespan. (A) Representative confocal images of whole midguts from control ( Luc RNAi ) and Tet RNAi adult flies driven by esg-Gal4, UAS-nGFP . Loss of ISCs/EBs (green) is restricted to the anterior midgut. (B) High-magnification images of the anterior and posterior midgut compartments across three independent Tet RNAi lines. (C) Quantification of ISC/EB percentage (GFP+/DAPI) in the anterior and posterior midgut. (D) Confirmation of ISC loss using the endogenous Tet-Gal4 T2A driver crossed with UAS-mCherry.nls and stained for the ISC marker Delta (green). (E) Quantification of ISC percentage (Delta+/DAPI) in the anterior midgut. (F) Survival curve of adult flies exposed to paraquat-induced oxidative stress over 48 hours. Tet RNAi n=54, Luciferase RNAi n=53. (G) Lifespan analysis comparing control and Tet RNAi adult flies. Tet RNAi #1 n=247, Tet RNAi #2 n=118, Luciferase RNAi n=240. Lifespans were analyzed with OASIS 2. Bars represent mean ± SD. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001. p values were from unpaired Student’s t-test.

Journal: bioRxiv

Article Title: Intrinsic regulation of intestinal stem cell fate and homeostasis by Tet

doi: 10.64898/2026.05.05.722984

Figure Lengend Snippet: Developmental depletion of Tet causes anterior-specific loss of ISCs and reduces adult lifespan. (A) Representative confocal images of whole midguts from control ( Luc RNAi ) and Tet RNAi adult flies driven by esg-Gal4, UAS-nGFP . Loss of ISCs/EBs (green) is restricted to the anterior midgut. (B) High-magnification images of the anterior and posterior midgut compartments across three independent Tet RNAi lines. (C) Quantification of ISC/EB percentage (GFP+/DAPI) in the anterior and posterior midgut. (D) Confirmation of ISC loss using the endogenous Tet-Gal4 T2A driver crossed with UAS-mCherry.nls and stained for the ISC marker Delta (green). (E) Quantification of ISC percentage (Delta+/DAPI) in the anterior midgut. (F) Survival curve of adult flies exposed to paraquat-induced oxidative stress over 48 hours. Tet RNAi n=54, Luciferase RNAi n=53. (G) Lifespan analysis comparing control and Tet RNAi adult flies. Tet RNAi #1 n=247, Tet RNAi #2 n=118, Luciferase RNAi n=240. Lifespans were analyzed with OASIS 2. Bars represent mean ± SD. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001. p values were from unpaired Student’s t-test.

Article Snippet: Additional lines included UAS - Tet (from Dahua Chen, Zheng et al. 2015), Tet-Gal4 T2A (BDSC #76666), 10xStatGFP (BDSC #26198), UAS-redstinger, UAS-Flp, Ubi-FRT.stop.GFP/cyo (G-TRACE, BDSC #28280), UAS-Luciferase RNAi (BDSC #31603), UAS-empty (from Hugo Bellen, Moulton et al. 2021), esgF/O ts (from Bruce Edgar, ), esg-Gal4, UAS-nGFP (from Lucy O’Brien), NRE-LacZ; esg-Gal4, UAS-nGFP and esg-Gal4, UAS-GFP; tub-Gal80ts (from Heinrich Jasper), esg-Gal4 (from Shigeo Hayashi, Hayashi et al. 2002), UAS-mCherry.nls (BDSC #38424), T2A-ATP8B (BDSC #91431), Cut-Gal4 (BDSC #27327), Ara-GFP (BDSC #93570), and T2A-Lerp (BDSC #77798).

Techniques: Control, Staining, Marker, Luciferase

Tet expression dynamically marks progenitors during midgut metamorphosis. (A) Overview of gut metamorphosis paired with representative images of Tet-Gal4 T2A > UAS-mCherry.nls expression from the larval (L3) stage through adulthood. (B) L3 midguts showing Tet expression (red) highly enriched in Peripheral Cells (PCs) marked by 10xStatGFP (green) and largely absent from AMPs. (C) Temporal dynamics of Tet expression (red) during early to late metamorphosis (3hr to 48hr APF), showing broad induction followed by selective retention in the developing adult gut. (D-F) High-magnification images at 72hr APF, 90hr APF, and adult stages demonstrating that Tet expression strictly co-localizes with the ISC/EBs. (G) Schematic summary model of AMP morphogenesis, fate diversification, and corresponding Tet expression dynamics. It is also worth noting that Tet is expressed outside the gut epithelium, in the surrounding visceral muscle.

Journal: bioRxiv

Article Title: Intrinsic regulation of intestinal stem cell fate and homeostasis by Tet

doi: 10.64898/2026.05.05.722984

Figure Lengend Snippet: Tet expression dynamically marks progenitors during midgut metamorphosis. (A) Overview of gut metamorphosis paired with representative images of Tet-Gal4 T2A > UAS-mCherry.nls expression from the larval (L3) stage through adulthood. (B) L3 midguts showing Tet expression (red) highly enriched in Peripheral Cells (PCs) marked by 10xStatGFP (green) and largely absent from AMPs. (C) Temporal dynamics of Tet expression (red) during early to late metamorphosis (3hr to 48hr APF), showing broad induction followed by selective retention in the developing adult gut. (D-F) High-magnification images at 72hr APF, 90hr APF, and adult stages demonstrating that Tet expression strictly co-localizes with the ISC/EBs. (G) Schematic summary model of AMP morphogenesis, fate diversification, and corresponding Tet expression dynamics. It is also worth noting that Tet is expressed outside the gut epithelium, in the surrounding visceral muscle.

Article Snippet: Additional lines included UAS - Tet (from Dahua Chen, Zheng et al. 2015), Tet-Gal4 T2A (BDSC #76666), 10xStatGFP (BDSC #26198), UAS-redstinger, UAS-Flp, Ubi-FRT.stop.GFP/cyo (G-TRACE, BDSC #28280), UAS-Luciferase RNAi (BDSC #31603), UAS-empty (from Hugo Bellen, Moulton et al. 2021), esgF/O ts (from Bruce Edgar, ), esg-Gal4, UAS-nGFP (from Lucy O’Brien), NRE-LacZ; esg-Gal4, UAS-nGFP and esg-Gal4, UAS-GFP; tub-Gal80ts (from Heinrich Jasper), esg-Gal4 (from Shigeo Hayashi, Hayashi et al. 2002), UAS-mCherry.nls (BDSC #38424), T2A-ATP8B (BDSC #91431), Cut-Gal4 (BDSC #27327), Ara-GFP (BDSC #93570), and T2A-Lerp (BDSC #77798).

Techniques: Expressing

Journal: bioRxiv

Article Title: Intrinsic regulation of intestinal stem cell fate and homeostasis by Tet

doi: 10.64898/2026.05.05.722984

Figure Lengend Snippet:

Article Snippet: Additional lines included UAS - Tet (from Dahua Chen, Zheng et al. 2015), Tet-Gal4 T2A (BDSC #76666), 10xStatGFP (BDSC #26198), UAS-redstinger, UAS-Flp, Ubi-FRT.stop.GFP/cyo (G-TRACE, BDSC #28280), UAS-Luciferase RNAi (BDSC #31603), UAS-empty (from Hugo Bellen, Moulton et al. 2021), esgF/O ts (from Bruce Edgar, ), esg-Gal4, UAS-nGFP (from Lucy O’Brien), NRE-LacZ; esg-Gal4, UAS-nGFP and esg-Gal4, UAS-GFP; tub-Gal80ts (from Heinrich Jasper), esg-Gal4 (from Shigeo Hayashi, Hayashi et al. 2002), UAS-mCherry.nls (BDSC #38424), T2A-ATP8B (BDSC #91431), Cut-Gal4 (BDSC #27327), Ara-GFP (BDSC #93570), and T2A-Lerp (BDSC #77798).

Techniques: In Vivo, Functional Assay, Quantitative Proteomics, Derivative Assay, Epifluorescence Microscopy