Journal: bioRxiv

Article Title: Metabolic control of enteroendocrine cell fate through a redox state sensor CtBP

doi: 10.1101/2025.06.30.662346

Figure Lengend Snippet: (A) ctbp deficiency ( CtBP 03463 /CtBP 87De-10 ) in Drosophila larvae results in a reduction on relative number of EE cells on HSD (30% sucrose), quantified in (B) . The data were compared with unpaired t-test, P = 0.0012. (C) Upper panel: The distribution of Dh31-, Tk-, and AstA-expressing EE cells in Drosophila larvae: a schematic overview. Lower panel: Whole-body ctbp loss-of-function ( CtBP 03463 /CtBP 87De-10 ) results in a loss of Dh31-, Tk-, and AstA-expressing EE cells on HSD. (D) Quantification of C. Two-tailed t-tests, *P < 0.01; **P = 0.0032. (E) Targeted loss of CtBP in EE cells (Pros-GAL4>CtBP RNAi) leads to reduced relative number of EE cells in larvae on HSD. Two-tailed t-test, P = 0.0046. (F) EE cell specific loss of CtBP function (Pros-GAL4 > CtBP RNAi ) on HSD reduces EE cell size (Mann-Whitney U test, P = 0.00313). (G) Loss of CtBP function in EE cells (Pros-GAL4>CtBP RNAi) leads to sugar intolerance of Drosophila larvae. The pupariation rate of larvae fed on HSD (20% sucrose) were compared with the log-rank test (χ = 88.6, P < 0.0001). (H) Intestinal lipid levels are reduced in larvae with EE-specific CtBP deficiency (Pros-GAL4>CtBP RNAi), quantified as relative Oil Red O intensity in (I) , unpaired t-test, **** P < 0.0001. (J) Loss of Dh31, Tk and AstA function in EE cells (Pros-GAL4> RNAi) leads to sugar intolerance of Drosophila larvae. The pupariation rate of larvae fed on HSD (20% sucrose) were compared using a log-rank test with the adjustment for multiple comparisons (χ = 59.4, P < 0.0001). ( K ) Intestinal lipid levels are reduced in larvae with EE-specific Dh31-deficiency (Pros-GAL4>Dh31 RNAi), quantified as relative Oil Red O intensity in ( L) , unpaired t-test, **** P < 0.0001.

Article Snippet: Following blocking, the tissues were incubated with antibodies against Prospero (1:500, Developmental Studies Hybridoma Bank, MR1A), Mesh (1:1000, a kind gift from Mikio Furuse), AstA (Developmental Studies Hybridoma Bank, 5F10C), Dh31 or Tk (a kind gift from Jan Veenstra).

Techniques: Expressing, Two Tailed Test, MANN-WHITNEY

Journal: bioRxiv

Article Title: Metabolic control of enteroendocrine cell fate through a redox state sensor CtBP

doi: 10.1101/2025.06.30.662346

Figure Lengend Snippet: (A) The ratiometric SoNar and iNap sensors were expressed by Pros-GAL4 ts driver to monitor the NAD + /NADH and NADPH/NADP + ratios in EE cells. GAL4 expression was activated 16 h prior EE cell redox state measurements. EE cells of HSD fed larvae showed elevated reduction of NAD + and NADP + in EE cells when compared to those of 2-deoxyglucose (2-DG)-fed animals. The data were analyzed using the Mann-Whitney test, with ****P < 0.0001. (B) Feeding larvae with nicotinamide riboside (NR), a precursor of nicotinamide adenine dinucleotide, increases the proportion of EE cells (Pros + ) in the Drosophila intestine. Data were analyzed using an unpaired t-test (****P < 0.0001). NR feeding also increases the number of Dh31-positive cells, with statistical significance determined by unpaired t-tests (*P = 0.0149; ***P = 0.0008). (C) EE cell specific knockdown (Pros-GAL4) of Naprt , a critical enzyme in NAD biosynthesis, leads to sugar intolerance. Pupariation rates of control and experimental genotype on HSD were compared using the log-rank test (χ = 64.2, P < 0.0001). (D) EE cell specific knockdown (Pros-GAL4) of Naprt reduces relative EE cell numbers in the Drosophila intestine as well as (E) number of Dh31, AstA and Tk positive cells. The data were analyzed using unpaired t-test, *P < 0.03; ****P < 0.0001. (F) Transgenes expressing the bacterial enzymes LbNOX and TPNOX, which oxidize NADH and NADPH in EE cells, respectively, impair larval development on HSD. Pupariation rate is analyzed using the log-rank test, revealing significant effects for both LbNOX (χ = 43.8, P < 0.0001) and TPNOX (χ = 54.1, P < 0.0001).

Article Snippet: Following blocking, the tissues were incubated with antibodies against Prospero (1:500, Developmental Studies Hybridoma Bank, MR1A), Mesh (1:1000, a kind gift from Mikio Furuse), AstA (Developmental Studies Hybridoma Bank, 5F10C), Dh31 or Tk (a kind gift from Jan Veenstra).

Techniques: Expressing, MANN-WHITNEY, Knockdown, Control

Journal: STAR Protocols

Article Title: Protocol for multiplexed RNAscope-based imaging of mRNAs in whole-mount adult Drosophila brains

doi: 10.1016/j.xpro.2025.103615

Figure Lengend Snippet:

Article Snippet: Mouse anti-AstA (1:100 dilution) , DSHB , 5F10-c.

Techniques: Recombinant, Plasmid Preparation, Multiplex Assay, RNAscope, Software, Microscopy