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a. Overview of an adult Drosophila gut with R1-R5 regions labelled. Scale bar, 100 microns. b, c Confocal images of trachea ( b ) and intestinal epithelium ( c ) from R4 and R5 regions of the posterior midgut in the context of Sucrose or Pe infection. Scale bar, 20 microns. d . Quantification of tracheal coverage in the R4 and R5 regions of the midgut upon Pe infection. (n=6,6,6,6). e . Quantification of cell aspect ratio in R4 and R5 regions of the gut upon infection. Each data point represents an individual enterocyte. (n =6,6,6,6). f . Tilescans across the R4 and R5 regions of the posterior midgut where enterocytes cell junctions have been detected with lgl (green). Scale bar, 50 microns. f’ . Representative image of inferred relative cell pressure calculated using <t>Bayesian</t> <t>Force</t> <t>Inference.</t> Cells of higher pressure are red/yellow and lower pressure cells blue/green. g . Quantification of relative pressure across the R4 (grey) and R5 (green) regions of the midgut upon sucrose feeding or Pe infection, measured by Bayesian Force Inference in MATLAB. Smaller circles represent individual cells and larger circles in foreground represent average of cells per gut (n=4,4,5,5). h . Representative image of nanoindentation setup. Drosophila gut is placed on cell-Tak adhesive and cantilever is used to detect stiffness by touching the outside of the gut. i . Quantification of stiffness across midgut R4 and R5 regions using nanoindentation in the context of sucrose feeding or Pe infection. Smaller circles represent individual measurements within a gut (n=4,5,5,3). j . Overview of GFP reconstitution across synaptic partners (GRASP) system. Components of a split GFP barrel are expressed in two different cell types where close contact results in GFP reconstitution and fluorescence. When expressed in the trachea ( btl-lexA ; red) and enterocytes ( Mex-gal4) GFP (green) reconstitution is observed upon Pe damage as a result of close contact. k . Confocal images of GRASP expression in the intestinal epithelium and surrounding trachea following sucrose or Pe infection. Scale bar, 20 microns. l . Quantification of the number of GRASP positive cells per posterior midgut (n=20,23). m, n . Controls without btl-lexA driver system ( m ) and without Mex-Gal4 driver system ( n ). Scale bar, 20 microns. Data represent the mean□±□s.e.m. n numbers indicated for each graph are listed from left to right and they correspond to the numbers of guts analysed. Statistics : l . Two-tailed unpaired Student’s t-test. d, e, g, i. Two-way ANOVA followed by Tukey’s multiple comparisons test *P=0.05, ** P = 0.01, *** P=0.001 and **** P= 0.0001. DAPI, 4,6-diamidino-2-phenylindole.
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a. Overview of an adult Drosophila gut with R1-R5 regions labelled. Scale bar, 100 microns. b, c Confocal images of trachea ( b ) and intestinal epithelium ( c ) from R4 and R5 regions of the posterior midgut in the context of Sucrose or Pe infection. Scale bar, 20 microns. d . Quantification of tracheal coverage in the R4 and R5 regions of the midgut upon Pe infection. (n=6,6,6,6). e . Quantification of cell aspect ratio in R4 and R5 regions of the gut upon infection. Each data point represents an individual enterocyte. (n =6,6,6,6). f . Tilescans across the R4 and R5 regions of the posterior midgut where enterocytes cell junctions have been detected with lgl (green). Scale bar, 50 microns. f’ . Representative image of inferred relative cell pressure calculated using <t>Bayesian</t> <t>Force</t> <t>Inference.</t> Cells of higher pressure are red/yellow and lower pressure cells blue/green. g . Quantification of relative pressure across the R4 (grey) and R5 (green) regions of the midgut upon sucrose feeding or Pe infection, measured by Bayesian Force Inference in MATLAB. Smaller circles represent individual cells and larger circles in foreground represent average of cells per gut (n=4,4,5,5). h . Representative image of nanoindentation setup. Drosophila gut is placed on cell-Tak adhesive and cantilever is used to detect stiffness by touching the outside of the gut. i . Quantification of stiffness across midgut R4 and R5 regions using nanoindentation in the context of sucrose feeding or Pe infection. Smaller circles represent individual measurements within a gut (n=4,5,5,3). j . Overview of GFP reconstitution across synaptic partners (GRASP) system. Components of a split GFP barrel are expressed in two different cell types where close contact results in GFP reconstitution and fluorescence. When expressed in the trachea ( btl-lexA ; red) and enterocytes ( Mex-gal4) GFP (green) reconstitution is observed upon Pe damage as a result of close contact. k . Confocal images of GRASP expression in the intestinal epithelium and surrounding trachea following sucrose or Pe infection. Scale bar, 20 microns. l . Quantification of the number of GRASP positive cells per posterior midgut (n=20,23). m, n . Controls without btl-lexA driver system ( m ) and without Mex-Gal4 driver system ( n ). Scale bar, 20 microns. Data represent the mean□±□s.e.m. n numbers indicated for each graph are listed from left to right and they correspond to the numbers of guts analysed. Statistics : l . Two-tailed unpaired Student’s t-test. d, e, g, i. Two-way ANOVA followed by Tukey’s multiple comparisons test *P=0.05, ** P = 0.01, *** P=0.001 and **** P= 0.0001. DAPI, 4,6-diamidino-2-phenylindole.
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a. Overview of an adult Drosophila gut with R1-R5 regions labelled. Scale bar, 100 microns. b, c Confocal images of trachea ( b ) and intestinal epithelium ( c ) from R4 and R5 regions of the posterior midgut in the context of Sucrose or Pe infection. Scale bar, 20 microns. d . Quantification of tracheal coverage in the R4 and R5 regions of the midgut upon Pe infection. (n=6,6,6,6). e . Quantification of cell aspect ratio in R4 and R5 regions of the gut upon infection. Each data point represents an individual enterocyte. (n =6,6,6,6). f . Tilescans across the R4 and R5 regions of the posterior midgut where enterocytes cell junctions have been detected with lgl (green). Scale bar, 50 microns. f’ . Representative image of inferred relative cell pressure calculated using <t>Bayesian</t> <t>Force</t> <t>Inference.</t> Cells of higher pressure are red/yellow and lower pressure cells blue/green. g . Quantification of relative pressure across the R4 (grey) and R5 (green) regions of the midgut upon sucrose feeding or Pe infection, measured by Bayesian Force Inference in MATLAB. Smaller circles represent individual cells and larger circles in foreground represent average of cells per gut (n=4,4,5,5). h . Representative image of nanoindentation setup. Drosophila gut is placed on cell-Tak adhesive and cantilever is used to detect stiffness by touching the outside of the gut. i . Quantification of stiffness across midgut R4 and R5 regions using nanoindentation in the context of sucrose feeding or Pe infection. Smaller circles represent individual measurements within a gut (n=4,5,5,3). j . Overview of GFP reconstitution across synaptic partners (GRASP) system. Components of a split GFP barrel are expressed in two different cell types where close contact results in GFP reconstitution and fluorescence. When expressed in the trachea ( btl-lexA ; red) and enterocytes ( Mex-gal4) GFP (green) reconstitution is observed upon Pe damage as a result of close contact. k . Confocal images of GRASP expression in the intestinal epithelium and surrounding trachea following sucrose or Pe infection. Scale bar, 20 microns. l . Quantification of the number of GRASP positive cells per posterior midgut (n=20,23). m, n . Controls without btl-lexA driver system ( m ) and without Mex-Gal4 driver system ( n ). Scale bar, 20 microns. Data represent the mean□±□s.e.m. n numbers indicated for each graph are listed from left to right and they correspond to the numbers of guts analysed. Statistics : l . Two-tailed unpaired Student’s t-test. d, e, g, i. Two-way ANOVA followed by Tukey’s multiple comparisons test *P=0.05, ** P = 0.01, *** P=0.001 and **** P= 0.0001. DAPI, 4,6-diamidino-2-phenylindole.
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a. Overview of an adult Drosophila gut with R1-R5 regions labelled. Scale bar, 100 microns. b, c Confocal images of trachea ( b ) and intestinal epithelium ( c ) from R4 and R5 regions of the posterior midgut in the context of Sucrose or Pe infection. Scale bar, 20 microns. d . Quantification of tracheal coverage in the R4 and R5 regions of the midgut upon Pe infection. (n=6,6,6,6). e . Quantification of cell aspect ratio in R4 and R5 regions of the gut upon infection. Each data point represents an individual enterocyte. (n =6,6,6,6). f . Tilescans across the R4 and R5 regions of the posterior midgut where enterocytes cell junctions have been detected with lgl (green). Scale bar, 50 microns. f’ . Representative image of inferred relative cell pressure calculated using Bayesian Force Inference. Cells of higher pressure are red/yellow and lower pressure cells blue/green. g . Quantification of relative pressure across the R4 (grey) and R5 (green) regions of the midgut upon sucrose feeding or Pe infection, measured by Bayesian Force Inference in MATLAB. Smaller circles represent individual cells and larger circles in foreground represent average of cells per gut (n=4,4,5,5). h . Representative image of nanoindentation setup. Drosophila gut is placed on cell-Tak adhesive and cantilever is used to detect stiffness by touching the outside of the gut. i . Quantification of stiffness across midgut R4 and R5 regions using nanoindentation in the context of sucrose feeding or Pe infection. Smaller circles represent individual measurements within a gut (n=4,5,5,3). j . Overview of GFP reconstitution across synaptic partners (GRASP) system. Components of a split GFP barrel are expressed in two different cell types where close contact results in GFP reconstitution and fluorescence. When expressed in the trachea ( btl-lexA ; red) and enterocytes ( Mex-gal4) GFP (green) reconstitution is observed upon Pe damage as a result of close contact. k . Confocal images of GRASP expression in the intestinal epithelium and surrounding trachea following sucrose or Pe infection. Scale bar, 20 microns. l . Quantification of the number of GRASP positive cells per posterior midgut (n=20,23). m, n . Controls without btl-lexA driver system ( m ) and without Mex-Gal4 driver system ( n ). Scale bar, 20 microns. Data represent the mean□±□s.e.m. n numbers indicated for each graph are listed from left to right and they correspond to the numbers of guts analysed. Statistics : l . Two-tailed unpaired Student’s t-test. d, e, g, i. Two-way ANOVA followed by Tukey’s multiple comparisons test *P=0.05, ** P = 0.01, *** P=0.001 and **** P= 0.0001. DAPI, 4,6-diamidino-2-phenylindole.

Journal: bioRxiv

Article Title: Intestinal Tissue Mechanics Regulate Angiogenesis and Stem Cell Proliferation via Vascular Piezo

doi: 10.1101/2025.04.16.649133

Figure Lengend Snippet: a. Overview of an adult Drosophila gut with R1-R5 regions labelled. Scale bar, 100 microns. b, c Confocal images of trachea ( b ) and intestinal epithelium ( c ) from R4 and R5 regions of the posterior midgut in the context of Sucrose or Pe infection. Scale bar, 20 microns. d . Quantification of tracheal coverage in the R4 and R5 regions of the midgut upon Pe infection. (n=6,6,6,6). e . Quantification of cell aspect ratio in R4 and R5 regions of the gut upon infection. Each data point represents an individual enterocyte. (n =6,6,6,6). f . Tilescans across the R4 and R5 regions of the posterior midgut where enterocytes cell junctions have been detected with lgl (green). Scale bar, 50 microns. f’ . Representative image of inferred relative cell pressure calculated using Bayesian Force Inference. Cells of higher pressure are red/yellow and lower pressure cells blue/green. g . Quantification of relative pressure across the R4 (grey) and R5 (green) regions of the midgut upon sucrose feeding or Pe infection, measured by Bayesian Force Inference in MATLAB. Smaller circles represent individual cells and larger circles in foreground represent average of cells per gut (n=4,4,5,5). h . Representative image of nanoindentation setup. Drosophila gut is placed on cell-Tak adhesive and cantilever is used to detect stiffness by touching the outside of the gut. i . Quantification of stiffness across midgut R4 and R5 regions using nanoindentation in the context of sucrose feeding or Pe infection. Smaller circles represent individual measurements within a gut (n=4,5,5,3). j . Overview of GFP reconstitution across synaptic partners (GRASP) system. Components of a split GFP barrel are expressed in two different cell types where close contact results in GFP reconstitution and fluorescence. When expressed in the trachea ( btl-lexA ; red) and enterocytes ( Mex-gal4) GFP (green) reconstitution is observed upon Pe damage as a result of close contact. k . Confocal images of GRASP expression in the intestinal epithelium and surrounding trachea following sucrose or Pe infection. Scale bar, 20 microns. l . Quantification of the number of GRASP positive cells per posterior midgut (n=20,23). m, n . Controls without btl-lexA driver system ( m ) and without Mex-Gal4 driver system ( n ). Scale bar, 20 microns. Data represent the mean□±□s.e.m. n numbers indicated for each graph are listed from left to right and they correspond to the numbers of guts analysed. Statistics : l . Two-tailed unpaired Student’s t-test. d, e, g, i. Two-way ANOVA followed by Tukey’s multiple comparisons test *P=0.05, ** P = 0.01, *** P=0.001 and **** P= 0.0001. DAPI, 4,6-diamidino-2-phenylindole.

Article Snippet: First, we used an available Bayesian Force Inference model in MATLAB to predict relative forces in the midgut tissue across the R4 and R5 regions from tile scan images of Pe infected and control guts, using the R5 gut region as internal control.

Techniques: Infection, Adhesive, Fluorescence, Expressing, Two Tailed Test