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(A) Schematic representation of the transgenes used to generate the control (Ac-G) and the acinar-derived pancreatic tumour model (Ac-K). (B-E) 5-day zebrafish larvae expressing the protein GFP ( B,D ) or GFP-KRAS G12D ( C,E ) in the acinar cells of the pancreas. White asterisk indicates gut autofluorescence. White arrows indicate the fluorescent heart, marked by the cmlc2:GFP reporter. Panels D and E show magnified views of the pancreata outlined with white dashed lines in panels B and C. i: principal islet, a: acinar cells. Scale bars : 200 µm. (F-G) Representative photographs of male zebrafish showing a marked abdominal protrusion, indicative of tumour presence ( G, white arrow), compared to a control male ( F ). Scale bars: 2mm. (H-I) Dissected digestive systems of an adult control zebrafish ( H ), showing a healthy pancreas (P, white dashes) or of an Ac-K p53 m/m fish ( I ) with a pancreatic tumour (T, yellow dashes). L: liver, G: gut, S: spleen, P: pancreas, T: tumour. Scale bars: 2mm. (J) Tumour incidence across various TP53 backgrounds. K-L: Hematoxylin and eosin (HE) staining of transversal sections of control pancreas (K) and Ac-K p53 m/m tumour (L) with close-up regions (K’,L’) outlined by black boxes. M-N : Immunofluorescence staining <t>with</t> <t>phospho-histone</t> <t>H3</t> <t>(pH3)</t> antibody on control (M ) and tumoral tissue sections (N). Control and tumoral pancreatic tissues are surrounded by green dashes. L; Liver, G; gut, d; extrapancreatic duct, i; principal islet, a; acinar cells. Scale bars: 200 µm. O : Quantification of pH3-positive cells in four Ac-K p53 m/m tumours compared to six control pancreas (3 p53 +/+ ; 1 p53 +/m , 2 p53 m/m ). The percentage of pH3 positive cells was calculated as the ratio of the pH3-positive area to the DAPI-positive area, based on the mean of two independent sections per fish. Each point represents an individual fish. Data are presented as mean ± SD; **P = 0.0095, determined using the Mann–Whitney test.
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(A) Schematic representation of the transgenes used to generate the control (Ac-G) and the acinar-derived pancreatic tumour model (Ac-K). (B-E) 5-day zebrafish larvae expressing the protein GFP ( B,D ) or GFP-KRAS G12D ( C,E ) in the acinar cells of the pancreas. White asterisk indicates gut autofluorescence. White arrows indicate the fluorescent heart, marked by the cmlc2:GFP reporter. Panels D and E show magnified views of the pancreata outlined with white dashed lines in panels B and C. i: principal islet, a: acinar cells. Scale bars : 200 µm. (F-G) Representative photographs of male zebrafish showing a marked abdominal protrusion, indicative of tumour presence ( G, white arrow), compared to a control male ( F ). Scale bars: 2mm. (H-I) Dissected digestive systems of an adult control zebrafish ( H ), showing a healthy pancreas (P, white dashes) or of an Ac-K p53 m/m fish ( I ) with a pancreatic tumour (T, yellow dashes). L: liver, G: gut, S: spleen, P: pancreas, T: tumour. Scale bars: 2mm. (J) Tumour incidence across various TP53 backgrounds. K-L: Hematoxylin and eosin (HE) staining of transversal sections of control pancreas (K) and Ac-K p53 m/m tumour (L) with close-up regions (K’,L’) outlined by black boxes. M-N : Immunofluorescence staining <t>with</t> <t>phospho-histone</t> <t>H3</t> <t>(pH3)</t> antibody on control (M ) and tumoral tissue sections (N). Control and tumoral pancreatic tissues are surrounded by green dashes. L; Liver, G; gut, d; extrapancreatic duct, i; principal islet, a; acinar cells. Scale bars: 200 µm. O : Quantification of pH3-positive cells in four Ac-K p53 m/m tumours compared to six control pancreas (3 p53 +/+ ; 1 p53 +/m , 2 p53 m/m ). The percentage of pH3 positive cells was calculated as the ratio of the pH3-positive area to the DAPI-positive area, based on the mean of two independent sections per fish. Each point represents an individual fish. Data are presented as mean ± SD; **P = 0.0095, determined using the Mann–Whitney test.
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(A) Schematic representation of the transgenes used to generate the control (Ac-G) and the acinar-derived pancreatic tumour model (Ac-K). (B-E) 5-day zebrafish larvae expressing the protein GFP ( B,D ) or GFP-KRAS G12D ( C,E ) in the acinar cells of the pancreas. White asterisk indicates gut autofluorescence. White arrows indicate the fluorescent heart, marked by the cmlc2:GFP reporter. Panels D and E show magnified views of the pancreata outlined with white dashed lines in panels B and C. i: principal islet, a: acinar cells. Scale bars : 200 µm. (F-G) Representative photographs of male zebrafish showing a marked abdominal protrusion, indicative of tumour presence ( G, white arrow), compared to a control male ( F ). Scale bars: 2mm. (H-I) Dissected digestive systems of an adult control zebrafish ( H ), showing a healthy pancreas (P, white dashes) or of an Ac-K p53 m/m fish ( I ) with a pancreatic tumour (T, yellow dashes). L: liver, G: gut, S: spleen, P: pancreas, T: tumour. Scale bars: 2mm. (J) Tumour incidence across various TP53 backgrounds. K-L: Hematoxylin and eosin (HE) staining of transversal sections of control pancreas (K) and Ac-K p53 m/m tumour (L) with close-up regions (K’,L’) outlined by black boxes. M-N : Immunofluorescence staining with phospho-histone H3 (pH3) antibody on control (M ) and tumoral tissue sections (N). Control and tumoral pancreatic tissues are surrounded by green dashes. L; Liver, G; gut, d; extrapancreatic duct, i; principal islet, a; acinar cells. Scale bars: 200 µm. O : Quantification of pH3-positive cells in four Ac-K p53 m/m tumours compared to six control pancreas (3 p53 +/+ ; 1 p53 +/m , 2 p53 m/m ). The percentage of pH3 positive cells was calculated as the ratio of the pH3-positive area to the DAPI-positive area, based on the mean of two independent sections per fish. Each point represents an individual fish. Data are presented as mean ± SD; **P = 0.0095, determined using the Mann–Whitney test.

Journal: bioRxiv

Article Title: Single-Cell Cross-Species Profiling identifies Conserved Transcriptional Networks in Early Pancreatic Tumourigenesis

doi: 10.64898/2026.03.03.708839

Figure Lengend Snippet: (A) Schematic representation of the transgenes used to generate the control (Ac-G) and the acinar-derived pancreatic tumour model (Ac-K). (B-E) 5-day zebrafish larvae expressing the protein GFP ( B,D ) or GFP-KRAS G12D ( C,E ) in the acinar cells of the pancreas. White asterisk indicates gut autofluorescence. White arrows indicate the fluorescent heart, marked by the cmlc2:GFP reporter. Panels D and E show magnified views of the pancreata outlined with white dashed lines in panels B and C. i: principal islet, a: acinar cells. Scale bars : 200 µm. (F-G) Representative photographs of male zebrafish showing a marked abdominal protrusion, indicative of tumour presence ( G, white arrow), compared to a control male ( F ). Scale bars: 2mm. (H-I) Dissected digestive systems of an adult control zebrafish ( H ), showing a healthy pancreas (P, white dashes) or of an Ac-K p53 m/m fish ( I ) with a pancreatic tumour (T, yellow dashes). L: liver, G: gut, S: spleen, P: pancreas, T: tumour. Scale bars: 2mm. (J) Tumour incidence across various TP53 backgrounds. K-L: Hematoxylin and eosin (HE) staining of transversal sections of control pancreas (K) and Ac-K p53 m/m tumour (L) with close-up regions (K’,L’) outlined by black boxes. M-N : Immunofluorescence staining with phospho-histone H3 (pH3) antibody on control (M ) and tumoral tissue sections (N). Control and tumoral pancreatic tissues are surrounded by green dashes. L; Liver, G; gut, d; extrapancreatic duct, i; principal islet, a; acinar cells. Scale bars: 200 µm. O : Quantification of pH3-positive cells in four Ac-K p53 m/m tumours compared to six control pancreas (3 p53 +/+ ; 1 p53 +/m , 2 p53 m/m ). The percentage of pH3 positive cells was calculated as the ratio of the pH3-positive area to the DAPI-positive area, based on the mean of two independent sections per fish. Each point represents an individual fish. Data are presented as mean ± SD; **P = 0.0095, determined using the Mann–Whitney test.

Article Snippet: Immunofluorescence staining was performed with anti-GFP (rabbit, Cell Signalling #2956S 1:200), anti-Caveolin-1(rabbit, Cell Signalling #3238 1:250), anti-α-Amylase (rabbit, Cell Signalling #3796 1:200), anti-Phospho-Histone H3 (pH3) antibodies (rabbit, Cell Signalling #3377T 1:200) overnight at 4°C followed by rabbit HRP SignalStain Boost (Cell Signalling #8114), and Tyramide-FITC, Tyramide-Cy3 (TSA Plus Cyanine 3 System #NEL744001KT Akoya BioSciences) or Tyramide-Cy5 substrate (TSA Plus Cyanine 5 System #NEL745001KT Akoya BioSciences).

Techniques: Control, Derivative Assay, Expressing, Staining, Immunofluorescence, MANN-WHITNEY